Moraxella(branhamella) catarrhalis antigens

ABSTRACT

The present invention relates to polypeptides of  Moraxella ( Branhamella )  calarrhalis  which may be useful for prophylaxis, diagnosis and/or therapy purposes.

FIELD OF THE INVENTION

[0001] The present invention is related to polypeptides, moreparticularly polypeptides of Moraxella (Branhamella) catarrhalis whichmay be used to prevent, diagnose and/or treat Moraxella (Branhamella)catarrhalis infection.

BACKGROUND OF THE INVENTION

[0002]Moraxella (Branhamella) catarrhalis is a Gram-negative diplococcusthat causes respiratory tract infections in humans. M. catarrhalis isnow accepted as the third most common cause of otitis media in infantsand children, after Streptococcus pneumoniae and Haemophilus influenzae.M. catarrhalis has also been associated with several other types ofinfection, including sinusitis, persistent cough, acute laryngitis inadults, suppurative keratitis, conjunctivitis neonatorum, and invasivediseases in the immunocompromised host.

[0003] Since approximately 90% of M. catarrhalis strains are resistantto antibiotics (β-lactamase positive) and that recurrent otitis media isassociated with high morbidity, there is a need for the development of avaccine that will protect hosts from M. catarrhalis infection. Aninfection by M. catarrhalis induces an immune response against antigensfound at the surface of the bacterial cells. However, many of thesesurface proteins are still not characterized, nor has the immuneresponse resulting in protection from infection by different strainsbeen determined.

[0004] To develop a vaccine that will protect hosts from M. catarrhalisinfection, efforts have mainly been concentrated on outer membraneproteins such as the high-molecular-mass protein named ubiquitoussurface protein A (UspA). This protein is considered a promising vaccinecandidate because a monoclonal antibody and polyclonal antibodies wereboth shown to be bactericidal and protective in the murinepulmonary-clearance model. However, this protein was shown to be highlyvariable among the different strains of M. catarrhalis. In addition tothis protein, other M. catarrhalis proteins have generated interest aspotential vaccine candidates. The transferrin-binding protein whichpossesses conserved epitopes exposed on the bacterial surface. However,there was divergence in the degree of antibody cross-reactivity with theprotein from one strain to another. Other investigators have alsofocused on the 45-kDa protein CD (OMP CD). This protein is highlyconserved among strains of M. catarrhalis, however adults with chronicobstructive pulmonary disease show variability in the immune responseagainst the OMP CD.

[0005] Therefore there remains an unmet need for M. catarrhalispolypeptides which may be used to prevent, diagnose and/or treatMoraxella (Branhamella) catarrhalis infection.

SUMMARY OF THE INVENTION

[0006] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 70%identity to a second polypeptide comprising a sequence chosen from SEQID Nos 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof.

[0007] According to one aspect, the present invention relates topolypeptides comprising a sequence chosen from SEQ ID No: 2, 4, 6, 8,10, 12, 14 or fragments or analogs thereof.

[0008] In other aspects, there are provided polypeptides encoded bypolynucleotides of the invention, pharmaceutical compositions, vectorscomprising polynucleotides of the invention operably linked to anexpression control region, as well as host cells transfected with saidvectors and processes for producing polypeptides comprising culturingsaid host cells under conditions suitable for expression.

BRIEF DESCRIPTION OF THE DRAWINGS

[0009]FIG. 1 represents the DNA sequence of BVH-MC2 gene from M.catarrhalis strain ETSU C-2; SEQ ID NO: 1. The underlined portion of thesequence represents the region coding for the leader peptide.

[0010]FIG. 2 represents the amino acid sequence of BVH-MC2 polypeptidefrom M. catarrhalis strain ETSU C-2; SEQ ID NO: 2. The underlinesequence represents the 30 amino acid residues leader peptide.

[0011]FIG. 3 represents the partial DNA sequence of BVH-MC2 gene from M.catarrhalis strain ETSU 658; SEQ ID NO: 3.

[0012]FIG. 4 represents the partial amino acid sequence of BVH-MC2polypeptide from M. catarrhalis strain ETSU 658; SEQ ID NO: 4.

[0013]FIG. 5 represents the partial DNA sequence of BVH-MC2 gene from M.catarrhalis strain ETSU T-25; SEQ ID NO: 5.

[0014]FIG. 6 represents the partial amino acid sequence of BVH-MC2polypeptide from M. catarrhalis strain ETSU T-25; SEQ ID NO: 6.

[0015]FIG. 7 represents the partial DNA sequence of BVH-MC2 gene from M.catarrhalis strain M-12; SEQ ID NO: 7.

[0016]FIG. 8 represents the partial amino acid sequence of BVH-MC2polypeptide from M. catarrhalis strain M-12; SEQ ID NO: 8.

[0017]FIG. 9 represents the DNA sequence of BVH-MC3 gene from M.catarrhalis strain ETSU C-2; SEQ ID NO: 9. The underlined portion of thesequence represents the region coding for the leader peptide.

[0018]FIG. 10 represents the amino acid sequence of BVH-MC3 polypeptidefrom M. catarrhalis strain ETSU C-2; SEQ ID NO: 10. The underlinesequence represents the 46 amino acid residues leader peptide.

[0019]FIG. 11 represents the DNA sequence of BVH-MC4 gene from M.catarrhalis strain ETSU C-2; SEQ ID NO: 11. The underlined portion ofthe sequence represents the region coding for the leader peptide.

[0020]FIG. 12 represents the amino acid sequence of BVH-MC4 polypeptideM. catarrhalis strain ETSU C-2; SEQ ID NO: 12. The underline sequencerepresents the 42 amino acid residues leader peptide.

[0021]FIG. 13 represents the DNA sequence of BVH-MCS gene from M.catarrhalis strain ETSU C-2; SEQ ID NO: 13. The underlined portion ofthe sequence represents the region coding for the leader peptide.

[0022]FIG. 14 represents the amino acid sequence of BVH-MC5 polypeptideM. catarrhalis strain ETSU C-2; SEQ ID NO: 14. The underline sequencerepresents the 60 amino acid residues leader peptide.

[0023]FIG. 15 depicts the comparison of the partial nucleotide sequencesof the BVH-MC2 genes from ETSU C-2, ETSU 658, ETSU T-25, and M-12 M.catarrhalis strains by using the program Clustal W from Macvectorsequence analysis software (version 6.5). Underneath the alignment,there is a consensus line where * and blank spaces respectivelyrepresent identical nucleotides and differences between sequences.

[0024]FIG. 16 depicts the comparison of the predicted amino acidsequences of the BVH-MC2 partial open reading frames from ETSU C-2, ETSU658, ETSU T-25, and M-12 M. catarrhalis strains by using the programClustal W from MacVector sequence analysis software (version 6.5).Underneath the alignment, there is a consensus line where * charactersrepresent identical amino acid residues.

DETAILED DESCRIPTION OF THE INVENTION

[0025] The present invention provides purified and isolatedpolynucleotides, which encode Moraxella polypeptides which may be usedto prevent, diagnose and/or treat Moraxella infection.

[0026] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 70%identity to a second polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof.

[0027] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 80%identity to a second polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof.

[0028] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 95%identity to a second polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof.

[0029] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 70%identity to a second polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12 or 14.

[0030] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 80%identity to a second polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12 or 14.

[0031] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 95%identity to a second polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12 or 14.

[0032] According to one aspect, the present invention relates topolypeptides comprising a sequence chosen from SEQ ID Nos: 2, 4, 6, 8,10, 12, 14 or fragments or analogs thereof.

[0033] According to one aspect, the present invention relates topolypeptides characterized by the amino acid sequence comprising SEQ IDNO: 2, 4, 6, 8, 10, 12 or 14 According to one aspect, the presentinvention provides a polynucleotide encoding an epitope bearing portionof a polypeptide comprising a sequence chosen from SEQ ID NO: 2, 4, 6,8, 10, 12, 14 or fragments or analogs thereof.

[0034] According to one aspect, the present invention provides apolynucleotide encoding an epitope bearing portion of a polypeptidecomprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10, 12 or 14.

[0035] According to one aspect, the present invention relates to epitopebearing portions of a polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof.

[0036] According to one aspect, the present invention relates to epitopebearing portions of a polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12 or 14.

[0037] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 70%identity to a second polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12 or 14.

[0038] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 80%identity to a second polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12 or 14.

[0039] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 90%identity to a second polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12 or 14.

[0040] According to one aspect, the present invention provides anisolated polynucleotide encoding a polypeptide having at least 95%identity to a second polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12 or 14.

[0041] According to one aspect, the present invention relates topolypeptides comprising a sequence chosen from SEQ ID No: 2, 4, 6, 8,10, 12 or 14.

[0042] According to one aspect, the present invention provides anisolated polynucleotide comprising a polynucleotide chosen from:

[0043] (a) a polynucleotide encoding a polypeptide having at least 70%identity to a second polypeptide comprising a sequence chosen from: SEQID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof;

[0044] (b) a polynucleotide encoding a polypeptide having at least 80%identity to a second polypeptide comprising a sequence chosen from: SEQID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof;

[0045] (c) a polynucleotide encoding a polypeptide having at least 95%identity to a second polypeptide comprising a sequence chosen from: SEQID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof;

[0046] (d) a polynucleotide encoding a polypeptide comprising a sequencechosen from: SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogsthereof;

[0047] (e) a polynucleotide encoding a polypeptide capable of generatingantibodies having binding specificity for a polypeptide comprising asequence chosen from: SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments oranalogs thereof;

[0048] (f) a polynucleotide encoding an epitope bearing portion of apolypeptide comprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10,12, 14 or fragments or analogs thereof;

[0049] (g) a polynucleotide comprising a sequence chosen from SEQ ID NO:1, 3, 5, 7, 9, 11, 13 or fragments or analogs thereof;

[0050] (h) a polynucleotide that is complementary to a polynucleotide in(a), (b), (c), (d), (e), (f) or (g).

[0051] According to one aspect, the present invention provides anisolated polynucleotide comprising a polynucleotide chosen from:

[0052] (a) a polynucleotide encoding a polypeptide having at least 70%identity to a second polypeptide comprising a sequence chosen from: SEQID NO: 2, 4, 6, 8, 10, 12 or 14;

[0053] (b) a polynucleotide encoding a polypeptide having at least 80%identity to a second polypeptide comprising a sequence chosen from: SEQID NO: 2, 4, 6, 8, 10, 12 or 14;

[0054] (c) a polynucleotide encoding a polypeptide having at least 95%identity to a second polypeptide comprising a sequence chosen from: SEQID NO: 2, 4, 6, 8, 10, 12 or 14;

[0055] (d) a polynucleotide encoding a polypeptide comprising a sequencechosen from: SEQ ID NO: 2, 4, 6, 8, 10, 12 or 14;

[0056] (e) a polynucleotide encoding a polypeptide capable of raisingantibodies having binding specificity for a polypeptide comprising asequence chosen from: SEQ ID NO: 2, 4, 6, 8, 10, 12 or 14;

[0057] (f) a polynucleotide encoding an epitope bearing portion of apolypeptide comprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10,12 or 14;

[0058] (g) a polynucleotide comprising a sequence chosen from SEQ ID NO:1, 3, 5, 7, 9, 11 and 13;

[0059] (h) a polynucleotide that is complementary to a polynucleotide in(a), (b), (c), (d), (e), (f) or (g).

[0060] According to one aspect, the present invention provides anisolated polypeptide comprising a polypeptide chosen from:

[0061] (a) a polypeptide having at least 70% identity to a secondpolypeptide comprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10,12, 14 or fragments or analogs thereof;

[0062] (b) a polypeptide having at least 80% identity to a secondpolypeptide comprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10,12, 14 or fragments or analogs thereof;

[0063] (c) a polypeptide having at least 95% identity to a secondpolypeptide comprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10,12, 14 or fragments or analogs thereof;

[0064] (d) a polypeptide comprising a sequence chosen from SEQ ID NO: 2,4, 6, 8, 10, 12, 14 or fragments or analogs thereof;

[0065] (e) a polypeptide capable of raising antibodies having bindingspecificity for a polypeptide comprising a sequence chosen from SEQ IDNO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof;

[0066] (f) an epitope bearing portion of a polypeptide comprising asequence chosen from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments oranalogs thereof;

[0067] (g) the polypeptide of (a), (b), (c), (d), (e) or (f) wherein theN-terminal Met residue is deleted;

[0068] (h) the polypeptide of (a), (b), (c), (d), (e) or (f) wherein thesecretory amino acid sequence is deleted.

[0069] According to one aspect, the present invention provides anisolated polypeptide comprising a polypeptide chosen from:

[0070] (a) a polypeptide having at least 70% identity to a secondpolypeptide comprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10,12 or 14;

[0071] (b) a polypeptide having at least 80% identity to a secondpolypeptide comprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10,12 or 14;

[0072] (c) a polypeptide having at least 95% identity to a secondpolypeptide comprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10,12 or 14;

[0073] (d) a polypeptide comprising a sequence chosen from SEQ ID NO: 2,4, 6, 8, 10, 12 or 14;

[0074] (e) a polypeptide capable of raising antibodies having bindingspecificity for a polypeptide comprising a sequence chosen from SEQ IDNO: 2, 4, 6, 8, 10, 12 or 14;

[0075] (f) an epitope bearing portion of a polypeptide comprising asequence chosen from SEQ ID NO: 2, 4, 6, 8, 10, 12 or 14;

[0076] (g) the polypeptide of (a), (b), (c), (d), (e) or (f) wherein theN-terminal Met residue is deleted;

[0077] (h) the polypeptide of (a), (b), (c), (d), (e) or (f) wherein thesecretory amino acid sequence is deleted.

[0078] Those skilled in the art will appreciate that the inventionincludes DNA molecules, i.e. polynucleotides and their complementarysequences that encode analogs such as mutants, variants, homologues andderivatives of such polypeptides, as described herein in the presentpatent application. The invention also includes RNA moleculescorresponding to the DNA molecules of the invention. In addition to theDNA and RNA molecules, the invention includes the correspondingpolypeptides and monospecific antibodies that specifically bind to suchpolypeptides.

[0079] In a further embodiment, the polypeptides in accordance with thepresent invention are antigenic.

[0080] In a further embodiment, the polypeptides in accordance with thepresent invention are immunogenic.

[0081] In a further embodiment, the polypeptides in accordance with thepresent invention can elicit an immune response in a host.

[0082] In a further embodiment, the present invention also relates topolypeptides which are able to raise antibodies having bindingspecificity to the polypeptides of the present invention as definedabove.

[0083] An antibody that “has binding specificity” is an antibody thatrecognizes and binds the selected polypeptide but which does notsubstantially recognize and bind other molecules in a sample, e.g., abiological sample, which naturally includes the selected peptide.Specific binding can be measured using an ELISA assay in which theselected polypeptide is used as an antigen.

[0084] In accordance with the present invention, “protection” in thebiological studies is defined by a significant increase in the survivalcurve, rate or period. Statistical analysis using the Log rank test tocompare survival curves, and Fisher exact test to compare survival ratesand numbers of days to death, respectively, might be useful to calculateP values and determine whether the difference between the two groups isstatistically significant. P values of 0.05 are regarded as notsignificant.

[0085] In an additional aspect of the invention there are providedantigenic/immunogenic fragments of the polypeptides of the invention, orof analogs thereof.

[0086] The fragments of the present invention should include one or moresuch epitopic regions or be sufficiently similar to such regions toretain their antigenic/immunogenic properties. Thus, for fragmentsaccording to the present invention the degree of identity is perhapsirrelevant, since they may be 100% identical to a particular part of apolypeptide or analog thereof as described herein. The present inventionfurther provides fragments having at least 10 contiguous amino acidresidues from the polypeptide sequences of the present invention. In oneembodiment, at least 15 contiguous amino acid residues. In oneembodiment, at least 20 contiguous amino acid residues.

[0087] The skilled person will appreciate that analogs of thepolypeptides of the invention will also find use in the context of thepresent invention, i.e. as antigenic/immunogenic material. Thus, forinstance proteins or polypeptides which include one or more additions,deletions, substitutions or the like are encompassed by the presentinvention.

[0088] As used herein, “fragments”, “analogs” or “derivatives” of thepolypeptides of the invention include those polypeptides in which one ormore of the amino acid residues are substituted with a conserved ornon-conserved amino acid residue (preferably conserved) and which may benatural or unnatural. In one embodiment, derivatives and analogs ofpolypeptides of the invention will have about 80% identity with thosesequences illustrated in the figures or fragments thereof. That is, 80%of the residues are the same. In a further embodiment, polypeptides willhave greater than 80% identity. In a further embodiment, polypeptideswill have greater than 85% identity. In a further embodiment,polypeptides will have greater than 90% identity. In a furtherembodiment, polypeptides will have greater than 95% identity. In afurther embodiment, polypeptides will have greater than 99% identity. Ina further embodiment, analogs of polypeptides of the invention will havefewer than about 20 amino acid residue substitutions, modifications ordeletions and more preferably less than 10.

[0089] These substitutions are those having a minimal influence on thesecondary structure and hydropathic nature of the polypeptide. Preferredsubstitutions are those known in the art as conserved, i.e. thesubstituted residues share physical or chemical properties such ashydrophobicity, size, charge or functional groups. These includesubstitutions such as those described by Dayhoff, M. in Atlas of ProteinSequence and Structure 5, 1978 and by Argos, P. in EMBO J. 8, 779-785,1989. For example, amino acids, either natural or unnatural, belongingto one of the following groups represent conservative changes: ala, pro,gly, gln, asn, ser, thr, val; cys, ser, tyr, thr; val, ile, leu, met,ala, phe; lys, arg, orn, his; and phe, tyr, trp, his.

[0090] The preferred substitutions also include substitutions ofD-enantiomers for the corresponding L-amino acids.

[0091] In an alternative approach, the analogs could be fusionpolypeptides, incorporating moieties which render purification easier,for example by effectively tagging the desired polypeptide. It may benecessary to remove the “tag” or it may be the case that the fusionpolypeptide itself retains sufficient antigenicity to be useful.

[0092] The percentage of homology is defined as the sum of thepercentage of identity plus the percentage of similarity or conservationof amino acid type.

[0093] In one embodiment, analogs of polypeptides of the invention willhave about 70% identity with those sequences illustrated in the figuresor fragments thereof. That is, 70% of the residues are the same. In afurther embodiment, polypeptides will have greater than 80% identity. Ina further embodiment, polypeptides will have greater than 85% identity.In a further embodiment, polypeptides will have greater than 90%identity. In a further embodiment, polypeptides will have greater than95% identity. In a further embodiment, polypeptides will have greaterthan 99% identity. In a further embodiment, analogs of polypeptides ofthe invention will have fewer than about 20 amino acid residuesubstitutions, modifications or deletions and more preferably less than10.

[0094] In one embodiment, analogs of polypeptides of the invention willhave about 70% homology with those sequences illustrated in the figuresor fragments thereof. In a further embodiment, polypeptides will havegreater than 80% homology. In a further embodiment, polypeptides willhave greater than 85% homology. In a further embodiment, polypeptideswill have greater than 90% homology. In a further embodiment,polypeptides will have greater than 95% homology. In a furtherembodiment, polypeptides will have greater than 99% homology. In afurther embodiment, analogs of polypeptides of the invention will havefewer than about 20 amino acid residue substitutions, modifications ordeletions and more preferably less than 10.

[0095] One can use a program such as the CLUSTAL program to compareamino acid sequences. This program compares amino acid sequences andfinds the optimal alignment by inserting spaces in either sequence asappropriate. It is possible to calculate amino acid identity or homologyfor an optimal alignment. A program like BLASTx will align the longeststretch of similar sequences and assign a value to the fit. It is thuspossible to obtain a comparison where several regions of similarity arefound, each having a different score. Both types of identity analysisare contemplated in the present invention.

[0096] In an alternative approach, the analogs or derivatives could befusion polypeptides, incorporating moieties which render purificationeasier, for example by effectively tagging the desired protein orpolypeptide, it may be necessary to remove the “tag” or it may be thecase that the fusion polypeptide itself retains sufficient antigenicityto be useful.

[0097] It is well known that it is possible to screen an antigenicpolypeptide to identify epitopic regions, i.e. those regions which areresponsible for the polypeptide's antigenicity or immunogenicity.Methods for carrying out such screening are well known in the art. Thus,the fragments of the present invention should include one or more suchepitopic regions or be sufficiently similar to such regions to retaintheir antigenic/immunogenic properties.

[0098] Thus, for fragments according to the present invention the degreeof identity is perhaps irrelevant, since they may be 100% identical to aparticular part of a polypeptide, analog as described herein.

[0099] Thus, what is important for analogs, derivatives and fragments isthat they possess at least a degree of the antigenicity/immunogenicityof the protein or polypeptide from which they are derived.

[0100] Also included are polypeptides which have fused thereto othercompounds which alter the polypeptides biological or pharmacologicalproperties i.e. polyethylene glycol (PEG) to increase half-life; leaderor secretory amino acid sequences for ease of purification; prepro- andpro-sequences; and (poly)saccharides.

[0101] Furthermore, in those situations where amino acid regions arefound to be polymorphic, it may be desirable to vary one or moreparticular amino acids to more effectively mimic the different epitopesof the different Moraxella strains.

[0102] Moreover, the polypeptides of the present invention can bemodified by terminal —NH₂ acylation (eg. by acetylation, or thioglycolicacid amidation, terminal carboxy amidation, e.g. with ammonia ormethylamine) to provide stability, increased hydrophobicity for linkingor binding to a support or other molecule.

[0103] Also contemplated are hetero and homo polypeptide multimers ofthe polypeptide fragments and analogues. These polymeric forms include,for example, one or more polypeptides that have been cross-linked withcross-linkers such as avidin/biotin, gluteraldehyde ordimethyl-superimidate. Such polymeric forms also include polypeptidescontaining two or more tandem or inverted contiguous sequences, producedfrom multicistronic mRNAs generated by recombinant DNA technology.

[0104] In a further embodiment, the present invention also relates tochimeric polypeptides which comprise one or more polypeptides orfragments or analogs thereof as defined in the figures of the presentapplication.

[0105] In a further embodiment, the present invention also relates tochimeric polypeptides comprising two or more polypeptides having asequence chosen from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments oranalogs thereof; provided that the polypeptides are linked as to formeda chimeric polypeptide.

[0106] In a further embodiment, the present invention also relates tochimeric polypeptides comprising two or more polypeptides comprising asequence chosen from SEQ ID NO: 2, 4, 6, 8, 10, 12 or 14 provided thatthe polypeptides are linked as to formed a chimeric polypeptide.

[0107] Preferably, a fragment, analog or derivative of a polypeptide ofthe invention will comprise at least one antigenic region i.e. at leastone epitope.

[0108] In order to achieve the formation of antigenic polymers (i.e.synthetic multimers), polypeptides may be utilized having bishaloacetylgroups, nitroarylhalides, or the like, where the reagents being specificfor thio groups. Therefore, the link between two mercapto groups of thedifferent polypeptides may be a single bond or may be composed of alinking group of at least two, typically at least four, and not morethan 16, but usually not more than about 14 carbon atoms.

[0109] In a particular embodiment, polypeptide fragments and analogs ofthe invention do not contain a starting residue, such as methionine(Met) or valine (Val). Preferably, polypeptides will not incorporate aleader or secretory sequence (signal sequence). The signal portion of apolypeptide of the invention may be determined according to establishedmolecular biological techniques. In general, the polypeptide of interestmay be isolated from a Moraxella culture and subsequently sequenced todetermine the initial residue of the mature protein and therefore thesequence of the mature polypeptide.

[0110] It is understood that polypeptides can be produced and/or usedwithout their start codon (methionine or valine) and/or without theirleader peptide to favor production and purification of recombinantpolypeptides. It is known that cloning genes without sequences encodingleader peptides will restrict the polypeptides to the cytoplasm of E.coli and will facilitate their recovery (Glick, B. R. and Pasternak, J.J. (1998) Manipulation of gene expression in prokaryotes. In “Molecularbiotechnology: Principles and applications of recombinant DNA”, 2ndedition, ASM Press, Washington D.C., p.109-143).

[0111] According to another aspect of the invention, there are alsoprovided (i) a composition of matter containing a polypeptide of theinvention, together with a carrier, diluent or adjuvant; (ii) apharmaceutical composition comprising a polypeptide of the invention anda carrier, diluent or adjuvant; (iii) a vaccine comprising a polypeptideof the invention and a carrier, diluent or adjuvant; (iv) a method forinducing an immune response against Moraxella, in a host, byadministering to the host, an immunogenically effective amount of apolypeptide of the invention to elicit an immune response, e.g., aprotective immune response to Moraxella; and particularly, (v) a methodfor preventing and/or treating a Moraxella infection, by administering aprophylactic or therapeutic amount of a polypeptide of the invention toa host in need.

[0112] According to another aspect of the invention, there are alsoprovided (i) a composition of matter containing a polynucleotide of theinvention, together with a carrier, diluent or adjuvant; (ii) apharmaceutical composition comprising a polynucleotide of the inventionand a carrier, diluent or adjuvant; (iii) a method for inducing animmune response against Moraxella, in a host, by administering to thehost, an immunogenically effective amount of a polynucleotide of theinvention to elicit an immune response, e.g., a protective immuneresponse to Moraxella; and particularly, (iv) a method for preventingand/or treating a Moraxella infection, by administering a prophylacticor therapeutic amount of a polynucleotide of the invention to a host inneed. Before immunization, the polypeptides of the invention can also becoupled or conjugated to carrier proteins such as tetanus toxin,diphtheria toxin, hepatitis B virus surface antigen, poliomyelitis virusVP1 antigen or any other viral or bacterial toxin or antigen or anysuitable proteins to stimulate the development of a stronger immuneresponse. This coupling or conjugation can be done chemically orgenetically. A more detailed description of peptide-carrier conjugationis available in Van Regenmortel, M. H. V., Briand J. P., Muller S.,Plaué S., <<Synthetic Polypeptides as antigens>> in LaboratoryTechniques in Biochemistry and Molecular Biology, Vol.19 (ed.) Burdou,R. H. & Van Knippenberg P. H. (1988), Elsevier New York.

[0113] According to another aspect, there are provided pharmaceuticalcompositions comprising one or more Moraxella polypeptides of theinvention in a mixture with a pharmaceutically acceptable adjuvant.Suitable adjuvants include (1) oil-in-water emulsion formulations suchas MF59™, SAF™, Ribi™; (2) Freund's complete or incomplete adjuvant; (3)salts i.e. AlK(SO₄)₂, AlNa(SO₄—)₂, AlNH₄(SO₄)₂, Al(OH) 3, AlPO₄, silica,kaolin; (4) saponin derivatives such as Stimulon™ or particles generatedtherefrom such as ISCOMs (immunostimulating complexes); (5) cytokinessuch as interleukins, interferons, macrophage colony stimulating factor(M-CSF), tumor necrosis factor (TNF); (6) other substances such ascarbon polynucleotides i.e. poly IC and poly AU, detoxified choleratoxin (CTB) and E. coli heat labile toxin for induction of mucosalimmunity. A more detailed description of adjuvant is available in areview by M. Z. I Khan et al. in Pharmaceutical Research, vol. 11, No. 1(1994) pp2-11, and also in another review by Gupta et al., in Vaccine,Vol. 13, No. 14, pp1263-1276 (1995) and in WO 99/24578. Preferredadjuvants include QuilA™, QS21™, Alhydrogel™ and Adjuphos™.

[0114] Pharmaceutical compositions of the invention may be administeredparenterally by injection, rapid infusion, nasopharyngeal absorption,dermoabsorption, or buccal or oral.

[0115] Pharmaceutical compositions of the invention are used for theprophylaxis of Moraxella infection and/or diseases and symptoms mediatedby Moraxella infection as described in Manual of Clinical Microbiology,P. R. Murray (Ed, in chief), E. J. Baron, M. A. Pfaller, F. C. Tenoverand R. H. Yolken. ASM Press, Washington, D.C. seventh edition, 1999,1773p. In one embodiment, pharmaceutical compositions of the presentinvention are used for the treatment or prophylaxis of otitis media,sinusitis, persistent cough, acute laryngitis, suppurative keratitis,conjunctivitis neonatorum. In one embodiment, vaccine compositions ofthe invention are used for the treatment or prophylaxis of Moraxellainfection and/or diseases and symptoms mediated by Moraxella infection.In a further embodiment, the Moraxella infection is Moraxellacatarrhalis.

[0116] In a further embodiment, the invention provides a method forprophylaxis or treatment of Moraxella infection in a host susceptible toMoraxella infection comprising administering to said host a prophylacticor therapeutic amount of a composition of the invention.

[0117] As used in the present application, the term “host” includesmammals. In a further embodiment, the mammal is human.

[0118] In a particular embodiment, pharmaceutical compositions areadministered to those hosts at risk of Moraxella infection such asneonates, infants, children, elderly and immunocompromised hosts.

[0119] In a particular embodiment, pharmaceutical compositions areadministered to those hosts at risk of Moraxella infection such asadults.

[0120] Pharmaceutical compositions are preferably in unit dosage form ofabout 0.001 to 100 μg/kg (antigen/body weight) and more preferably 0.01to 10 μg/kg and most preferably 0.1 to 1 μg/kg 1 to 3 times with aninterval of about 1 to 6 week intervals between immunizations.

[0121] Pharmaceutical compositions are preferably in unit dosage form ofabout 0.1 μg to 10 mg and more preferably 1 g to 1 mg and mostpreferably 10 to 100 μg 1 to 3 times with an interval of about 1 to 6week intervals between immunizations.

[0122] According to another aspect, there are provided polynucleotidesencoding polypeptides characterized by the amino acid sequencecomprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 orfragments or analogs thereof. In one embodiment, polynucleotides arethose illustrated in SEQ ID No: 1, 3, 5, 7, 9, 11, 13 which may includethe open reading frames (ORF), encoding the polypeptides of theinvention.

[0123] It will be appreciated that the polynucleotide sequencesillustrated in the figures may be altered with degenerate codons yetstill encode the polypeptides of the invention. Accordingly the presentinvention further provides polynucleotides which hybridize to thepolynucleotide sequences herein above described (or the complementsequences thereof) having 70% identity between sequences. In oneembodiment, at least 80% identity between sequences. In one embodiment,at least 85% identity between sequences. In one embodiment, at least 90%identity between sequences. In a further embodiment, polynucleotides arehybridizable under stringent conditions i.e. having at least 95%identity. In a further embodiment, more than 97% identity.

[0124] Suitable stringent conditions for hybridation can be readilydetermined by one of skilled in the art (see for example Sambrook etal., (1989) Molecular cloning: A Laboratory Manual, 2^(nd) ed, ColdSpring Harbor, N.Y.; Current Protocols in Molecular Biology, (1999)Edited by Ausubel F. M. et al., John Wiley & Sons, Inc., N.Y.).

[0125] In a further embodiment, the present invention providespolynucleotides that hybridize under stringent conditions to either

[0126] (a) a DNA sequence encoding a polypeptide or

[0127] (b) the complement of a DNA sequence encoding a polypeptide;

[0128] wherein said polypeptide comprises a sequence chosen from SEQ IDNO: 2, 4, 6, 8, 10, 12 or 14 or fragments or analogs thereof.

[0129] In a further embodiment, the present invention providespolynucleotides that hybridize under stringent conditions to either

[0130] (a) a DNA sequence encoding a polypeptide or

[0131] (b) the complement of a DNA sequence encoding a polypeptide;

[0132] wherein said polypeptide comprises a sequence chosen from SEQ IDNO: 2, 4, 6, 8, 10, 12 or 14.

[0133] In a further embodiment, the present invention providespolynucleotides that hybridize under stringent conditions to either

[0134] (a) a DNA sequence encoding a polypeptide or

[0135] (b) the complement of a DNA sequence encoding a polypeptide;

[0136] wherein said polypeptide comprises at least 10 contiguous aminoacid residues from a polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12 or 14 or fragments or analogs thereof.

[0137] In a further embodiment, the present invention providespolynucleotides that hybridize under stringent conditions to either

[0138] (a) a DNA sequence encoding a polypeptide or

[0139] (b) the complement of a DNA sequence encoding a polypeptide;

[0140] wherein said polypeptide comprises at least 10 contiguous aminoacid residues from a polypeptide comprising a sequence chosen from SEQID NO: 2, 4, 6, 8, 10, 12 or 14.

[0141] In a further embodiment, polynucleotides are those illustrated inSEQ ID NO: 1, 3, 5, 7, 9, 11, 13 or fragments or analogs thereofencoding polypeptides of the invention.

[0142] In a further embodiment, polynucleotides are those illustrated inSEQ ID NO: 1, 3, 5, 7, 9, 11 or 13 encoding polypeptides of theinvention. As will be readily appreciated by one skilled in the art,polynucleotides include both DNA and RNA.

[0143] The present invention also includes polynucleotides complementaryto the polynucleotides described in the present application.

[0144] In a further aspect, polynucleotides encoding polypeptides of theinvention, or fragments, analogs or derivatives thereof, may be used ina DNA immunization method. That is, they can be incorporated into avector which is replicable and expressible upon injection therebyproducing the antigenic polypeptide in vivo. For example polynucleotidesmay be incorporated into a plasmid vector under the control of the CMVpromoter which is functional in eukaryotic cells. Preferably the vectoris injected intramuscularly.

[0145] According to another aspect, there is provided a process forproducing polypeptides of the invention by recombinant techniques byexpressing a polynucleotide encoding said polypeptide in a host cell andrecovering the expressed polypeptide product. Alternatively, thepolypeptides can be produced according to established synthetic chemicaltechniques i.e. solution phase or solid phase synthesis of oligopeptideswhich are ligated to produce the full polypeptide (block ligation)

[0146] General methods for obtention and evaluation of polynucleotidesand polypeptides are described in the following references: Sambrook etal, Molecular Cloning: A Laboratory Manual, 2nd ed, Cold Spring Harbor,N.Y., 1989; Current Protocols in Molecular Biology, Edited by Ausubel F.M. et al., John Wiley and Sons, Inc. New York; PCR Cloning Protocols,from Molecular Cloning to Genetic Engineering, Edited by White B. A.,Humana Press, Totowa, N.J., 1997, 490 pages; Protein Purification,Principles and Practices, Scopes R. K.; Springer-verlag, New York, 3rdEdition, 1993, 380 pages; Current Protocols in Immunology, Edited byColigan J. E. et al., John Wiley & Sons Inc., New York.

[0147] The present invention provides a process for producing apolypeptide comprising culturing a host cell of the invention underconditions suitable for expression of said polypeptide.

[0148] For recombinant production, host cells are transfected withvectors which encode the polypeptides of the invention, and thencultured in a nutrient media modified as appropriate for activatingpromoters, selecting transformants or amplifying the genes. Suitablevectors are those that are viable and replicable in the chosen host andinclude chromosomal, non-chromosomal and synthetic DNA sequences e.g.bacterial plasmids, phage DNA, baculovirus, yeast plasmids, vectorsderived from combinations of plasmids and phage DNA. The polypeptidesequence may be incorporated in the vector at the appropriate site usingrestriction enzymes such that it is operably linked to an expressioncontrol region comprising a promoter, ribosome binding site (consensusregion or Shine-Dalgarno sequence), and optionally an operator (controlelement). One can select individual components of the expression controlregion that are appropriate for a given host and vector according toestablished molecular biology principles (Sambrook et al, MolecularCloning: A Laboratory Manual, 2nd ed, Cold Spring Harbor, N.Y., 1989;Current Protocols in Molecular Biology, Edited by Ausubel F. M. et al.,John Wiley and Sons, Inc. New York). Suitable promoters include but arenot limited to LTR or SV40 promoter, E. coli lac, tac or trp promotersand the phage lambda PL promoter. Vectors will preferably incorporate anorigin of replication as well as selection markers i.e. ampicilinresistance gene. Suitable bacterial vectors include pET, pQE70, pQE60,pQE-9, pD10 phagescript, psiX174, pbluescript SK, pbsks, pNH8A, pNH16a,pNH18A, pNH46A, ptrc99a, pKK223-3, pKK233-3, pDR540, pRIT5 andeukaryotic vectors pBlueBacIII, pWLNEO, pSV2CAT, pOG44, pXT1, pSG,pSVK3, pBPV, pMSG and pSVL. Host cells may be bacterial i.e. E. coli,Bacillus subtilis, Streptomyces; fungal i.e. Aspergillus niger,Aspergillus nidulins; yeast i.e. Saccharomyces or eukaryotic i.e. CHO,COS.

[0149] Upon expression of the polypeptide in culture, cells aretypically harvested by centrifugation then disrupted by physical orchemical means (if the expressed polypeptide is not secreted into themedia) and the resulting crude extract retained to isolate thepolypeptide of interest. Purification of the polypeptide from culturemedia or lysate may be achieved by established techniques depending onthe properties of the polypeptide i.e. using ammonium sulfate or ethanolprecipitation, acid extraction, anion or cation exchange chromatography,phosphocellulose chromatography, hydrophobic interaction chromatography,hydroxylapatite chromatography and lectin chromatography. Finalpurification may be achieved using HPLC.

[0150] The polypeptides may be expressed with or without a leader orsecretion sequence. In the former case the leader may be removed usingpost-translational processing (see U.S. Pat. No. 4,431,739; U.S. Pat.No. 4,425,437; and U.S. Pat. No. 4,338,397) or be chemically removedsubsequent to purifying the expressed polypeptide.

[0151] According to a further aspect, the Moraxella polypeptides of theinvention may be used in a diagnostic test for Moraxella infection, inparticular Moraxella catarrhalis infection. Several diagnostic methodsare possible, for example detecting Moraxella organism in a biologicalsample, the following procedure may be followed:

[0152] a) obtaining a biological sample from a host;

[0153] b) incubating an antibody or fragment thereof reactive with aMoraxella polypeptide of the invention with the biological sample toform a mixture; and

[0154] c) detecting specifically bound antibody or bound fragment in themixture which indicates the presence of Moraxella.

[0155] Alternatively, a method for the detection of antibody specific toa Moraxella antigen in a biological sample containing or suspected ofcontaining said antibody may be performed as follows:

[0156] a) obtaining a biological sample from a host;

[0157] b) incubating one or more Moraxella polypeptides of the inventionor fragments thereof with the biological sample to form a mixture; and

[0158] c) detecting specifically bound antigen or bound fragment in themixture which indicates the presence of antibody specific to Moraxella.

[0159] One of skill in the art will recognize that this diagnostic testmay take several forms, including an immunological test such as anenzyme-linked immunosorbent assay (ELISA), a radioimmunoassay or a latexagglutination assay, essentially to determine whether antibodiesspecific for the protein are present in an organism.

[0160] The DNA sequences encoding polypeptides of the invention may alsobe used to design DNA probes for use in detecting the presence ofMoraxella in a biological sample suspected of containing such bacteria.The detection method of this invention comprises:

[0161] a) obtaining the biological sample from a host;

[0162] b) incubating one or more DNA probes having a DNA sequenceencoding a polypeptide of the invention or fragments thereof with thebiological sample to form a mixture; and

[0163] c) detecting specifically bound DNA probe in the mixture whichindicates the presence of Moraxella bacteria.

[0164] The DNA probes of this invention may also be used for detectingcirculating Moraxella i.e. Moraxella nucleic acids in a sample, forexample using a polymerase chain reaction, as a method of diagnosingMoraxella infections.

[0165] The probe may be synthesized using conventional techniques andmay be immobilized on a solid phase, or may be labelled with adetectable label. A preferred DNA probe for this application is anoligomer having a sequence complementary to at least about 6 contiguousnucleotides of the Moraxella polypeptides of the invention. In a furtherembodiment, the preferred DNA probe will be an oligomer having asequence complementary to at least about 15 contiguous nucleotides ofthe Moraxella polypeptides of the invention. In a further embodiment,the preferred DNA probe will be an oligomer having a sequencecomplementary to at least about 30 contiguous nucleotides of theMoraxella polypeptides of the invention. In a further embodiment, thepreferred DNA probe will be an oligomer having a sequence complementaryto at least about 50 contiguous nucleotides of the Moraxellapolypeptides of the invention.

[0166] Another diagnostic method for the detection of Moraxella in ahost comprises:

[0167] a) labelling an antibody reactive with a polypeptide of theinvention or fragment thereof with a detectable label;

[0168] b) administering the labelled antibody or labelled fragment tothe host; and

[0169] c) detecting specifically bound labelled antibody or labelledfragment in the host which indicates the presence of Moraxella.

[0170] Alternatively, a method for the detection of antibody specific toa Moraxella antigen in a biological sample containing or suspected ofcontaining said antibody may be performed as follows:

[0171] a) obtaining a biological sample from a host;

[0172] b) incubating one or more Moraxella polypeptides of the inventionor fragments thereof with the biological sample to form a mixture; and

[0173] c) detecting specifically bound antigen or bound fragment in themixture which indicates the presence of antibody specific to Moraxella.

[0174] One of skill in the art will recognize that the diagnostic testmay take several forms, including an immunological test such as anenzyme-linked immunosorbent assay (ELISA), a radioimmunoassay or a latexagglutination assay, essentially to determine whether antibodiesspecific for the protein are present in an organism.

[0175] The DNA sequences encoding polypeptides of the invention may alsobe used to design DNA probes for use in detecting the presence ofMoraxella in a biological sample suspected of containing such bacteria.The detection method of this invention comprises:

[0176] a) obtaining the biological sample from a host;

[0177] b) incubating one or more DNA probes having a DNA sequenceencoding a polypeptide of the invention or fragments thereof with thebiological sample to form a mixture; and

[0178] c) detecting specifically bound DNA probe in the mixture whichindicates the presence of Moraxella bacteria.

[0179] According to one aspect, the present invention provides the useof an antibody for treatment and/or prophylaxis of Moraxella infections.

[0180] A further aspect of the invention is the use of the Moraxellapolypeptides of the invention as immunogens for the production ofspecific antibodies for the diagnosis and in particular the treatment ofMoraxella infection. Suitable antibodies may be determined usingappropriate screening methods, for example by measuring the ability of aparticular antibody to passively protect against Moraxella infection ina test model. One example of an animal model is the mouse modeldescribed in the examples herein. The antibody may be a whole antibodyor an antigen-binding fragment thereof and may belong to anyimmunoglobulin class. The antibody or fragment may be of animal origin,specifically of mammalian origin and more specifically of murine, rat orhuman origin. It may be a natural antibody or a fragment thereof, or ifdesired, a recombinant antibody or antibody fragment. The termrecombinant antibody or antibody fragment means antibody or antibodyfragment which was produced using molecular biology techniques. Theantibody or antibody fragments may be polyclonal, or preferablymonoclonal. It may be specific for a number of epitopes associated withthe Moraxella polypeptides but is preferably specific for one.

[0181] According to one aspect, the present invention provides the useof an antibody for prophylaxis and/or treatment of Moraxella infections.

[0182] A further aspect of the invention is the use of the Moraxellapolypeptides of the invention as immunogens for the production ofspecific antibodies for the diagnosis and in particular the treatment ofMoraxella infection. Suitable antibodies may be determined usingappropriate screening methods, for example by measuring the ability of aparticular antibody to passively protect against Moraxella infection ina test model. One example of an animal model is the mouse modeldescribed in the examples herein. The antibody may be a whole antibodyor an antigen-binding fragment thereof and may belong to anyimmunoglobulin class. The antibody or fragment may be of animal origin,specifically of mammalian origin and more specifically of murine, rat orhuman origin. It may be a natural antibody or a fragment thereof, or ifdesired, a recombinant antibody or antibody fragment. The termrecombinant antibody or antibody fragment means antibody or antibodyfragment which was produced using molecular biology techniques. Theantibody or antibody fragments may be polyclonal, or preferablymonoclonal. It may be specific for a number of epitopes associated withthe Moraxella polypeptides but is preferably specific for one.

[0183] A further aspect of the invention is the use of the antibodiesdirected to the polypeptides of the invention for passive immunization.One could use the antibodies described in the present application.

[0184] A further aspect of the invention is a method for immunization,whereby an antibody raised by a polypeptide of the invention isadministered to a host in an amount sufficient to provide a passiveimmunization.

[0185] In a further embodiment, the invention provides the use of apharmaceutical composition of the invention in the manufacture of amedicament for the prophylactic or therapeutic treatment of Moraxellainfection.

[0186] In a further embodiment, the invention provides a kit comprisinga polypeptide of the invention for detection or diagnosis of Moraxellainfection.

[0187] Unless otherwise defined, all technical and scientific terms usedherein have the same meaning as commonly understood by one of ordinaryskill in the art to which this invention belongs. All publications,patent applications, patents, and other references mentioned herein areincorporated by reference in their entirety. In case of conflict, thepresent specification, including definitions, will control. In addition,the materials, methods, and examples are illustrative only and notintended to be limiting.

EXAMPLE 1

[0188] This example illustrates the cloning and molecularcharacteristics of BVH-MC2 gene and corresponding polypeptide.

[0189] The coding region of M. catarrhalis BVH-MC2 (SEQ ID NO: 1) genewas amplified by PCR (DNA Thermal Cycler GeneAmp PCR system 2400 PerkinElmer, San Jose, Calif.) from genomic DNA of M. catarrhalis strain ETSUC-2 using the following oligos that contained base extensions for theAddition of restriction sites NdeI (CATATG) and XhoI (CTCGAG): DMAR544(5′-CATCAGTGCATATGAATACGACACACCATCACACG-3′); DMAR545(5′-GAGTTATTCTCGAGTTTGTCCAAATTTGGCTTAGTTTTAC-3′). PCR products werepurified from agarose gel using a QIAquick gel extraction kit fromQIAgen following the manufacturer's instructions (Chatsworth, Calif.),and digested with NdeI and XhoI (Amersham Pharmacia Biotech, Inc, Baied'Urfe, Canada). The pET21b(+) vector (Novagen, Madison, Wis.) wasdigested with NdeI and XhoI and purified from agarose gel using aQIAquick gel extraction kit from QIAgen (Chatsworth, Calif.). TheNdeI-XhoI PCR products were ligated to the NdeI-XhoI pET21b(+)expression vector. The ligated products were transformed into E. colistrain DH5α [φ80dlacZΔM15 Δ(lacZYA-argF)U169 endA1 recA1hsdR17(r_(K)-m_(K)+) deOR thi-1 supE44 λ⁻gyrA96 relA1 (Gibco BRL,Gaithersburg, Md.) according to the method of Simanis (Hanahan, D. DNACloning, 1985, D. M. Glover (ed), pp. 109-135). Recombinant pET21b(+)plasmid (rpET21b(+)) containing BVH-MC2 gene was purified using a QIAgenkit (Chatsworth, Calif.) and DNA insert was sequenced (Taq Dye DeoxyTerminator Cycle Sequencing kit, ABI, Foster City, Calif.). TABLE 1Oligonucleotide primers used for PCR amplification of M. catarrhalisgenes. Primers Restriction Genes I.D. site Vector Sequence BVH-MC2DMAR544 NdeI pET21b 5′-CATCAGTGCATATGAATACGACACACCATCACACG-3′ (+) (SEQID No: 15) BVH-MC2 DMAR545 XhoI pET21b5′-GAGTTATTCTCGAGTTTGTCCAAATTTGGCTTAGTTTTAC-3′ (+) (SEQ ID No: 16)BVH-MC2 DMAR544a BglII pCMV-GH 5′-TCAGTGAGATCTTGAATACGACACACCATC-3′ (SEQID No: 17) BVH-MC2 DMAR545a SalI pCMV-GH5′-GATTTGAGTTGTCGACTTATTTGTCCAAATTTG-3′ (SEQ ID No: 18) BVH-MC3 DMAR592NdeI pET21b 5′-CGGAGTGCCATATGAGCTTAATTAATAAATTAAATG-3′ (+) (SEQ ID No:19) BVH-MC3 BMAR593 XhoI pET21b 5′-TATAACTCGAGGTTTTGTGCAACAGGTGTTG-3′(+) (SEQ ID No: 20) BVH-MC3 DMAR592a BglII pCMV-GH5′-CGCTTGAGATCTTGGAAGATGTGTATCAGCGTGC-3′ (SEQ ID No: 21) BVH-MC3DMAR593a HindIII pCMV-GH 5′-CAATAACAAAGCTTTCAGTTTTGTGCAACAGGTGTTG-3′(SEQ ID No: 22) BVH-MC4 RIOS71 NdeI pET21b5′-AACCGCACATATGTATCGCTTGGTGTCACCACC-3′ (+) (SEQ ID No: 23) BVH-MC4RIOS72 XhoI pET21b 5′-GGTGACTCGAGGTACTCATCACCAACTAATCGCAC-3′ (+) (SEQ IDNo: 24) BVH-MC4 RIOS71a BamHI pCMV-GH 5′-GCAGGATCCTTATCGCTTGGTGTCACC-3′(SEQ ID No: 25) BVH-MC4 RIOS72a SalI pCMV-GH5′-ATCAATCGGGTCGACTTAGTACTCATCACCA-3′ (SEQ ID No: 26) BVH-MC5 RIOS59NdeI pET21b 5′-AAAGCTTCATATGGCCCAAAGTCAAGAATCTGCC-3′ (+) (SEQ ID No: 27)BVH-MC5 RIOS60 XhoI pET21b 5′-CGATAACTCGAGTTGAACATCAGGCACCTGC-3′ (+)(SEQ ID No: 28) BVH-MC5 RIOS59a BglII pCMV-GH5′-ACCATTCAAAAGAGATCTTGGCCCAAAGTCAAGAATCTG-3′ (SEQ ID No: 29) BVH-MC5RIOS60a SalI pCMV-GH 5′-GTTAGACCGAGTCGACTCATTGAACATCAGGCA-3′ (SEQ ID No:30)

[0190] It was determined that the open reading frame (ORF) which codesfor BVH-MC2 polypeptide contains 1467-bp and encodes a 488 amino acidresidues polypeptide with a predicted pI of 6.08 and a predictedmolecular mass of 53754.35 Da. Analysis of the predicted amino acidresidues sequence (SEQ ID NO: 2) using the Spscan sofware (WisconsinSequence Analysis Package; Genetics Computer Group) suggested theexistence of a 30 amino acid residues signal peptide(MDTDMKHLTKHRLSAAIIGVLLFISPSVQA), which ends with a cleavage sitelocated between an alanine and an asparagine residues.

[0191] To confirm the presence by PCR amplification of BVH-MC2 (SEQ IDNO: 1) gene, the following 4 distinct M. catarrhalis strains were used:M. catarrhalis ETSU C-2, ETSU T-25, and ETSU 658 clinical isolates wereprovided by the East Tennessee State University; M. catarrhalis strainM-12 was provided by the centre de recherche en infectiologie du centrehospitalier de l'université Laval. The E. coli XL1-Blue MRF′ was used inthese experiments as negative control. BVH-MC2 (SEQ ID NO: 1) gene wasamplified by PCR (DNA Thermal Cycler GeneAmp PCR-system 2400 PerkinElmer, San Jose, Calif.) from genomic DNA from the 4 M. catarrhalisstrains, and the control E. coli strain using the oligonucleotidesprimers DMAR544 and DMAR545 (Table 1). PCR was performed with 30 cyclesof 30 sec at 94° C., 30 sec at 51° C. and 1 min 20 sec at 72° C. and afinal elongation period of 7 min at 72° C. The PCR products were sizefractionated in 1% agarose gels and were visualized by ethidium bromidestaining. The results of these PCR amplifications are presented in Table2. The analysis of the amplification products revealed that BVH-MC2 (SEQID NO: 1) gene was present in the genome of all of the 4 M. catarrhalisstrains tested. No such product was detected when the control E. coliDNA was submitted to identical PCR amplifications with theseoligonucleotide primers.

[0192] Sequencing of additional BVH-MC2 genes from other strainsconfirmed the high level of molecular conservation of this gene among M.catarrhalis strains. The respective coding region of M. catarrhalisBVH-MC2 gene from strains ETSU 658 (SEQ ID NO: 3), ETSU T-25 (SEQ ID NO:5)., and M-12 (SEQ ID NO: 7) were amplified by PCR using theoligonucleotide primers DMAR544 and DMAR545 as described above. PCRproducts were purified from agarose gel using a QIAquick gel extractionkit from QIAgen following the manufacturer's instructions (Chatsworth,Calif.) and the DNA insert were sequenced (Taq Dye Deoxy TerminatorCycle Sequencing kit, ABI, Foster City, Calif.). The total sequencecould be obtained in the same manner as in Example 1. The predictedamino acid sequences from strains ETSU C-2 (SEQ ID NO: 2), ETSU 658 (SEQID NO: 4), ETSU T-25 (SEQ ID NO: 6), and M-12 (SEQ ID NO: 8) wererespectively presented in the following FIGS. 2, 4, 6, and 8. The FIGS.15 and 16 respectively depict the consensus nucleotide and predictedamino acid sequences established for M. catarrhalis BVH-MC2. Pairwisecomparison of the BVH-MC2 predicted polypeptide sequences revealed 100%identity. This latter result clearly demonstrates the high level ofmolecular conservation of BVH-MC2 polypeptides among M. catarrhalisisolates. TABLE 2 Identification of M. catarrhalis genes by PCRamplification. Strain Identification by PCR amplification ofIdentification BVH-MC2 BVH-MC3 BVH-MC4 BVH-MC5 ETSU C-2 + + + + ETSU658 + + + + ETSU T-25 + + + + M-12 + + + + E. coil − − − −

EXAMPLE 2

[0193] This example illustrates the cloning and molecularcharacteristics of BVH-MC3 gene and corresponding polypeptide.

[0194] The coding region of M. catarrhalis BVH-MC3 (SEQ ID NO: 9) genewas amplified by PCR (DNA Thermal Cycler GeneAmp PCR system 2400 PerkinElmer, San Jose, Calif.) from genomic DNA of M. catarrhalis strain ETSUC-2 using the following oligos that contained base extensions for theaddition of restriction sites NdeI (CATATG) and XhoI (CTCGAG): DMARS92and DMAR593, which are presented in Table 1. The methods used forcloning BVH-MC3 into an expression vector and sequencing are similar tothe methods described in Example 1.

[0195] It was determined that the open reading frame (ORF) which codesfor BVH-MC3 contains 1656-bp and encodes a 551 amino acid residuespolypeptide with a predicted pI of 4.68 and a predicted molecular massof 58910.13 Da. Analysis of the predicted amino acid residues sequence(SEQ ID NO: 10) using the Spscan sofware (Wisconsin Sequence AnalysisPackage; Genetics Computer Group) suggested the existence of a 46 aminoacid residues signal peptide (MSLINKLNERITPHVLTSIKNQDGDNADKSNLLTAFYTIFAGRLSN), which ends with a cleavage sitelocated between an asparigine and a glutamic acid residues.

[0196] The BVH-MC3 gene was shown to be present after PCR amplificationusing the oligonucleotide primers DMAR592 and DMAR593 in the 4 M.catarrhalis strains tested (Table 2). The methods used for PCRamplification of the BVH-MC3 gene were similar to the methods presentedin Example 1. No such product was detected when the control E. coli DNAwas submitted to identical PCR amplification with these oligonucleotideprimers.

EXAMPLE 3

[0197] This example illustrates the cloning and molecularcharacteristics of BVH-MC4 gene and corresponding polypeptide.

[0198] The coding region of M. catarrhalis BVH-MC4 (SEQ ID NO: 11) genewas amplified by PCR (DNA Thermal Cycler GeneAmp PCR system 2400 PerkinElmer, San Jose, Calif.) from genomic DNA of M. catarrhalis strain ETSUC-2 using the following oligos that contained base extensions for theaddition of restriction sites NdeI (CATATG) and XhoI (CTCGAG): RIOS71and RIOS72, which are presented in Table 1. The methods used for cloningBVH-MC4 into an expression vector and sequencing are similar to themethods described in Example 1.

[0199] It was determined that the open reading frame —(ORF) which codesfor BVH-MC4 contains 125l-bp and encodes a 416 amino acid residuespolypeptide with a predicted pI of 4.84 and a predicted molecular massof 46125.11 Da. Analysis of the predicted amino acid residues sequence(SEQ ID NO: 12) using the Spscan sofware (Wisconsin Sequence AnalysisPackage; Genetics Computer Group) suggested the existence of a 42 aminoacid residues signal peptide(MDTKHIQQNWLLPDGVADVLFTDAQKQESLRDALLFVLTAHG), which ends with a cleavagesite located between an glycine and a tyrosine residues.

[0200] The BVH-MC4 gene was shown to be present after PCR amplificationusing the oligonucleotide primers RIOS71 and RIOS72 in the 4 M.catarrhalis strains tested (Table 2). The methods used for PCRamplification of the BVH-MC4 gene were similar to the methods presentedin Example 1. No such product was detected when the control E. coli DNAwas submitted to identical PCR amplification with these oligonucleotideprimers.

EXAMPLE 4

[0201] This example illustrates the cloning and molecularcharacteristics of BVH-MC5 gene and corresponding polypeptide.

[0202] The coding region of M. catarrhalis BVH-MCS (SEQ ID NO: 13) genewas amplified by PCR (DNA Thermal Cycler GeneAmp PCR system 2400 PerkinElmer, San Jose, Calif.) from genomic DNA of M. catarrhalis strain ETSUC-2 using the following oligos that contained base extensions for theaddition of restriction sites NdeI (CATATG) and XhoI (CTCGAG): RIOS59and RIOS60, which are presented in Table 1. The methods used for cloningBVH-MC5 into an expression vector and sequencing are similar to themethods described in Example 1.

[0203] It was determined that the open reading frame (ORF) which codesfor BVH-MC5 contains 639-bp and encodes a 212 amino acid residuespolypeptide with a predicted pI of 7.45 and a predicted molecular massof 24020.08 Da. Analysis of the predicted amino acid residues sequence(SEQ ID NO: 14) using the Spscan sofware (Wisconsin Sequence AnalysisPackage; Genetics Computer Group) suggested the existence of a 60 aminoacid residues signal peptide(MNNFVYQLQSFWYELNQVNRHTIAQSPKYIQLTVLGLIVMIIGIFGWLLAIL PTIQKLNA), whichends with a cleavage site located between two alanine residues.

[0204] The BVH-MC5 gene was shown to be present after PCR amplificationusing the oligonucleotide primers RIOS59 and RIOS60 in the 4 M.catarrhalis strains tested (Table 2). The methods used for PCRamplification of the BVH-MC5 gene were similar to the methods presentedin Example 1. No such product was detected when the control E. coli DNAwas submitted to identical PCR amplification with these oligonucleotideprimers.

EXAMPLE 5

[0205] This example illustrates the cloning of M. catarrhalis genes inCMV plasmid pCMV-GH.

[0206] The DNA coding regions of a M. catarrhalis polypeptides wereinserted in phase downstream of a human growth hormone (hGH) gene whichwas under the transcriptional control of the cytomegalovirus (CMV)promotor in the plasmid vector pCMV-GH (Tang et al., Nature, 1992,356:152). The CMV promotor is non-functional plasmid in E. coli cellsbut active upon administration of the plasmid in eukaryotic cells. Thevector also incorporated the ampicillin resistance gene.

[0207] The coding regions of BVH-MC2 (SEQ ID NO: 1), BVH-MC3 (SEQ ID NO:9), BVH-MC4 (SEQ ID NO: 11), and BVH-MC5 (SEQ ID NO: 13) genes withouttheir leader peptide regions were amplified by PCR (DNA Thermal CyclerGeneAmp PCR system 2400 Perkin Elmer, San Jose, Calif.) from genomic DNAof M. catarrhalis strain ETSU C-2 using oligonucleotide primers thatcontained base extensions for the addition of restriction sites BamHI(GGATCC), BglII (AGATCT), SalI (GTCGAC), or HindIII (AAGCTT) which aredescribed in Table 1. The PCR products were purified from agarose gelusing a QIAquick gel extraction kit from QIAgen (Chatsworth, Calif.),digested with restriction enzymes (Amersham Pharmacia Biotech, Inc, Baied'Urfe, Canada). The pCMV-GH vector (Laboratory of Dr. Stephen A.Johnston, Department of Biochemistry, The University of Texas, Dallas,Tex.) was digested with BamHI, BglII, SalI, or HindIII and purified fromagarose gel using the QIAquick gel extraction kit from QIAgen(Chatsworth, Calif.). The digested DNA fragments were ligated to thedigested pCMV-GH vector to create the hGH-BVH-MC2, hGH-BVH-MC3,hGH-BVH-MC4, and hGH-BVH-MC5 fusion polypeptides under the control ofthe CMV promoter. The ligated products were transformed into E. colistrain DH5a ([φ80dlacZΔM15 Δ(lacZYA-argF)U169 endA1 recA1hsdR17(r_(K)-m_(K)+) deOR thi-1 supE44 λ⁻gyrA96 relA1] (Gibco BRL,Gaithersburg, Md.) according to the method of Simanis (Hanahan, D. DNACloning, 1985, D. M. Glover (ed), pp. 109-135). The recombinant pCMVplasmids were purified using a QIAgen kit (Chatsworth, Calif.) and thenucleotides sequence of the DNA inserts were verified by DNA sequencing.

EXAMPLE 6

[0208] This example illustrates the use of DNA to elicit an immuneresponse to M. catarrhalis polypeptide antigens.

[0209] A group of 8 female BALB/c mice (Charles River, St-Constant,Québec, Canada) were immunized by intramuscular injection of 100 μlthree times at two- or three-week intervals with 50 μg of recombinantpCMV-GH encoding BVH-MC2 (SEQ ID NO: 1), BVH-MC3 (SEQ ID NO: 9), BVH-MC4(SEQ ID NO: 11), and BVH-MC5 (SEQ ID NO: 13) genes in presence of 50 μgof granulocyte-macrophage colony-stimulating factor (GM-CSF)-expressingplasmid pCMV-GH-GM-CSF (Laboratory of Dr. Stephen A. Johnston,Department of Biochemistry, The University of Texas, Dallas, Tex.). Ascontrol, a group of mice were injected with 50 μg of pCMV-GH in presenceof 50 μg of pCMV-GH-GM-CSF. Blood samples were collected from theorbital sinus prior to each immunization and seven days following thethird injection and serum antibody responses were determined by ELISAusing the corresponding His-Tag labeled M. catarrhalis recombinantpolypeptides as coating antigen. The production and purification ofthese His-tagged labeled M. catarrhalis recombinant polypeptides arepresented in Example 7.

EXAMPLE 7

[0210] This example illustrates the production and purification of M.catarrhalis recombinant polypeptides.

[0211] The recombinant pET21b(+) plasmid with BVH-MC2 (SEQ ID NO: 1),BVH-MC3 (SEQ ID NO: 9), BVH-MC4 (SEQ ID NO: 11), and BVH-MC5 (SEQ ID NO:13) genes were used to transform by electroporation (Gene Pulser IIapparatus, BIO-RAD Labs, Mississauga, Canada) E. coli strain AD494 (DE3)[Δara-leu7697 ΔlacX74 ΔphoA PvuII phOR ΔmalF3 F′ lac⁺ (lacI^(q)) pro]trxB::Kan (DE3)] (Novagen, Madison, Wis.). In this strain of E. coli,the T7 promotor controlling expression of the recombinant polypeptide isspecifically recognized by the T7 RNA polymerase (present on the λDE3prophage) whose gene is under the control of the lac promotor which isinducible by isopropyl-β-d-thio-galactopyranoside (IPTG). Thetransformant AD494(DE3)/rpET21b(+) was grown at 37° C. with agitation at250 rpm in LB broth (peptone log/L, yeast extract 5 g/L, NaCl 10 g/L)containing 100 μg of carbenicillin (Sigma-Aldrich Canada Ltd., Oakville,Canada) per ml until the A₆₀₀ reached a value of 0.5. In order to inducethe production of His-tagged M. catarrhalis recombinant polypeptides,the cells were incubated for 3 additional hours in the presence of IPTGat a final concentration of 1 mM. Induced cells from a 500 ml culturewere pelleted by centrifugation and frozen at −70° C.

[0212] The purification of the recombinant polypeptides from the solublecytoplasmic fraction of IPTG-induced AD494(DE3)/rpET21b(+) was done byaffinity chromatography based on the properties of the His*Tag sequence(6 consecutive histidine residues) to bind to divalent cations (Ni²⁺)immobilized on the His*Bind metal chelation resin. Briefly, the pelletedcells obtained from a 500 mL culture induced with IPTG was resuspendedin lysis buffer (20 mM Tris, 500 mM NaCl, 10 mM imidazole, pH 7.9)containing 1 mM PMSF, sonicated and centrifuged at 12,000×g for 20 minto remove debris. The supernatant was deposited on a Ni-NTA agarosecolumn (Qiagen, Mississauga, Ontario, Canada). The His-tagged labeled M.catarrhalis recombinant polypeptides were eluted with 250 mMimidazole-500 mM NaCl-20 mM Tris pH 7.9. The removal of the salt andimidazole from the sample was done by dialysis against PBS at 4° C. Thequantities of recombinant polypeptides obtained from the solublefraction of E. coli was estimated by MicroBCA (Pierce, Rockford, Ill.).

EXAMPLE 8

[0213] This example illustrates the reactivity of the His-tagged M.catarrhalis recombinant polypeptides with human palatine tonsils andsera collected from mice after immunization with M. catarrhalisantigenic preparations.

[0214] As shown in Table 3, BVH-MC2, BVH-MC3, and BVH-MC4 His-taggedrecombinant polypeptides were recognized in immunoblots by theantibodies present in the human palatine tonsils. It indicates thathumans, which are normally in contact with M. catarrhalis do developantibodies that are specific to these polypeptides. These particularhuman antibodies might be implicated in the protection against M.catarrhalis infection. In addition, immunoblots also revealed that seracollected from mice immunized with M. catarrhalis antigenic preparationenriched membrane polypeptides which induced significant lung clearancein a mouse model also developed antibodies that recognized BVH-MC2His-tagged recombinant polypeptides. These results indicate that thispolypeptide was present in M. catarrhalis antigenic preparation thatprotected mice against infection and that it induced antibodies thatreacted with the corresponding BVH-MC2 His-tagged recombinantpolypeptide. TABLE 3 Reactivity in immunoblots of human palatine tonsilsand sera collected from mice after immunization with M. catarrhalisantigenic preparations with M. catarrhalis His-tagged fusion recombinantpolypeptides. Purified Apparent Reactivity in immunoblots withrecombinant molecular weight Human palatine polypeptide I.D.¹ (kDa)²tonsils³ Mouse sera⁴ BVH-MC2 50 + − BVH-MC3 70 + + BVH-MC4 40 + −BVH-MC5 20 − −

EXAMPLE 9

[0215] This example illustrates the accessibility to antibodies of theBVH-MC2, BVH-MC3, BVH-MC4, and BVH-MC5 polypeptides at the surface of M.catarrhalis strain.

[0216] Bacteria were grown in Brain Heart Infusion (BHI) brothcontaining 0.25% dextrose at 37° C. in a 8% CO₂ atmosphere to give anOD_(490 nm) of 0.650 (˜10⁸ CFU/ml) Dilutions of anti-BVH-MC2,anti-BVH-MC3, anti-BVH-MC4, anti-BVH-MC5, or control sera were thenadded and allowed to bind to the cells, which were incubated for 2 h at4° C. with rotation. Samples were washed 4 times in blocking buffer[phosphate-buffered saline (PBS) containing 2% bovine serum albumin(BSA)], and then 1 ml of goat fluorescein (FITC)-conjugated anti-mouseIgG Fc (gamma) fragment specific diluted in blocking buffer was added.After an additional incubation of 60 min at room temperature withrotation in the dark, samples were washed 4 times in blocking buffer andfixed with 0.25% formaldehyde in PBS buffer for 18 h at 4° C. Cells werewashed 2 times in PBS buffer and resuspended in 0.5 ml of PBS buffer.Cells were kept in the dark at 4° C. until analyzed by flow cytometry(Epics® XL; Beckman Coulter, Inc.). Flow cytometric analysis revealedthat BVH-MC2-, BVH-MC3-, BVH-MC4-, and BVH-MC5-specific antibodiesefficiently recognized their corresponding surface exposed epitopes onthe homologous (ETSU C-2) M. catarrhalis strain tested (Table 4). It wasdetermined that more than 70% of the 10,000 Moraxella cells analyzedwere labeled with the antibodies present in the specific sera. Theseobservations clearly demonstrate that these polypeptides are accessibleat the surface where they can be easily recognized by antibodies.Anti-M. catarrhalis antibodies were shown to play an important role inthe protection against M. catarrhalis infection. TABLE 4 Evaluation ofthe attachment of BVH-MC2-, BVH-MC3-, BVH-MC4-, and BVH-MC5-specificantibodies at the surface of intact cells of M. catarrhalis strainETSU-C2. Serum Identification Fluorescence Index² % of labeled cells³Pool of BVH-MC2-specific 3.6 72.8 sera¹ Pool of BVH-MC3-specific 7.582.8 sera Pool of BVH-MC4-specific 10.9 92.4 sera Pool ofBVH-MC5-specific 6.7 77.4 sera Pool of negative control 1 7.4 sera⁴Positive control serum⁵ 43.8 98.7

EXAMPLE 10

[0217] This example illustrates the bactericidal activities ofanti-BVH-MC2 mouse sera.

[0218] Bacteria were plated on chocolate agar plate and incubated at 37°C. in a 8% CO₂ atmosphere for 18 h. Bacterial cells were thenresuspended in bacteriolysis buffer [10% Hanks' Balanced Salt Solution(HBSS) and 1% hydrolyzed casein, pH 7.3] to an OD_(490nm) of 0.25 anddiluted to 8×CFU/ml. The bactericidal assay was performed by mixing 25μl of the bacterial suspension with 50 μl of diluted heat-inactivatedtest serum and 15 μl of HBSS and incubating for 15 min at 37° C., 8% CO₂with agitation (200 rpm). The rabbit complement-containing serum wasthen added to a final concentration of 10%, and the mixture wasincubated for an additional 60 min at 37° C., 8% CO₂ with agitation (200rpm) At the end of the incubation period, the number of viable bacteriawas determined by plating 10 μl of the assay mixture on chocolate agarplate. The plates were incubated at 37° C. in an 8% CO₂ atmosphere for18-24 h. The control consisted of bacteria incubated withheat-inactivated sera collected from mice before immunization and rabbitcomplement.

[0219] The % of lysis was determined by the following mathematicalformula: $100 - \left\lbrack {\frac{A}{B} \times 100} \right\rbrack$

[0220] A=CFU obtained when the bacteria were incubated with immune sera

[0221] B=CFU obtained with pre-bleed sera

[0222] The M. catarrhalis strain ETSU 658 was used to evaluate thebactericidal activity of the sera. Percentage of lysis of 71.3 wasdetermined for mouse sera collected after immunization with purifiedrecombinant BVH-MC2 polypeptide (SEQ ID NO: 2) (Table 5). TABLE 5Evaluation of the bactericidal activities of anti- BVH-MC2 mouse sera.Identification Bactericidal titer % of lysis Pool of BVH-MC2- 1/35 71.3specific sera¹ Positive control 1/35 92.7 serum²

EXAMPLE 11

[0223] This example illustrates the bactericidal activities ofanti-BVH-MC3 mouse sera.

[0224] Bacteria were plated on chocolate agar plates and incubated at37° C. in a 8% CO₂ atmosphere for 18 h. Bacterial ells were thenresuspended in bacteriolysis buffer [10% Hanks' Balanced Salt Solution(HBSS) and 1% hydrolyzed casein, pH 7.3] to an OD_(490nm) of 0.25 anddiluted to 8×10⁴ CFU/ml. The bactericidal assay was performed by mixing25 μl of the bacterial suspension with 50 μl of diluted heat-inactivatedtest serum and 15 μl of HBSS and incubating for 15 min at 37° C., 8% CO₂with agitation (200 rpm). The rabbit complement-containing serum wasthen added to a final concentration of 10%, and the mixture wasincubated for an additional 60 min at 37° C., 8% CO₂ with agitation (200rpm) At the end of the incubation period, the number of viable bacteriawas determined by plating 101 of the assay mixture on chocolate agarplate. The plates were incubated at 37° C. in an 8% CO₂ atmosphere for18-24 h. The control consisted of bacteria incubated withheat-inactivated sera collected from mice before immunization and rabbitcomplement. The M. catarrhalis strain ETSU 658 was used to evaluate thebactericidal activity of the sera. The % of lysis was determined by thefollowing mathematical formula:$100 - \left\lbrack {\frac{A}{B} \times 100} \right\rbrack$

[0225] A=CFU obtained when the bacteria were incubated with immune sera

[0226] B=CFU obtained with pre-bleed sera

[0227] Bactericidal antibodies were found to be present in the seracollected from the 7 mice that were immunized with the purifiedrecombinant BVH-MC3 polypeptide (Table 6). No bactericidal activity wererecorded in the sera collected from control mice (data not shown). TABLE6 Evaluation of the bactericidal activity of anti- BVE-MC3 mouse sera.Serum identification¹ % of lysis S1² 33.3 S2 67.9 S3 89.6 S4 66.2 S578.0 S6 90.1 S7 37.1 Positive control serum² 77.3

EXAMPLE 12

[0228] This example illustrates the protection of mice against M.catarrhalis infection induced by immunization with purified recombinantBVH-MC3 polypeptide.

[0229] Groups of female BALB/c mice (Charles River) were immunizedsubcutaneously five times at two-week intervals with 20 μg of affinitypurified His-tagged M. catarrhalis recombinant BVH-MC3 polypeptide inpresence of 10% of QuilA adjuvant (Cedarlane Laboratories Ltd, Hornby,Canada) or, as control, with QuilA adjuvant alone in PBS. Blood sampleswere collected from the orbital sinus on day 0, 14, 28, 42, and 56 priorto each immunization and 14 days (day 70) following the fifth injection.One week later the mice were challenged intrapulmonary withapproximately 1×10⁶ CFU of the M. catarrhalis strain ETSU 658. Samplesof the M. catarrhalis challenge inoculum were plated on chocolate agarplates to determine the CFU and to verify the challenge dose. Mice werekilled by an intraperitoneal injection of sodium pentobarbital(Euthanyl™) 5 h after infection. The intact lungs were excised andhomogenised in a tissue homogeniser. The lung homogenate were assessedfor bacterial clearance by plating the serial dilutions for CFUdetermination. The % of clearance was determined by the followingmathematical formula:$100 - \left\lbrack {\frac{A}{B} \times 100} \right\rbrack$

[0230] A=CFU obtained with the mice immunized with the BVH-MC3polypeptide

[0231] B=CFU obtained in the control mice

[0232] As shown in Table 7, a reduction of 54% in the number of livingbacteria were determined for mice immunized with BVH-MC3 polypeptidecomparatively to mice in the control group. Thus, immunization withrecombinant BVH-MC3 polypeptide promoted rapid clearance of anheterologous strain of M. catarrhalis from the lungs of the mice. TABLE7 Pulmonary clearance of Moraxella catarrhalis by mice immunized withpurified recombinant BVH-MC3 polypeptide Bacterial recovery Bacterialrecovery from control group from BVH-MC3 group Bacterial (CFU/ml of lung(CFU/ml of lung clearance homogenate)^(a) homogenate)^(b) (%)^(c) 2.4 ×10⁵ ± 1.9 × 10⁵ 1.1 × 10⁵ ± 7.9 × 10⁴ 54

[0233]

1 30 1 1467 DNA Moraxella catarrhalis 1 atggacactg acatgaaaca tttaacaaaacatcgcctat cagctgccat cattggcgtt 60 ttattattca ttagcccatc agtgcaagcaaatacgacac accatcacac gctaaccagt 120 agcgagctta aacttgctga tgatagtattattgatagta tcaatcaatt gggtgagctg 180 accgtcaata ttccaaatac acaatattttcaaaccaaca acggtgtgag cgttgctttt 240 acgccattac atgagctgcc tattgtcgatatcagcttgt attttaatgc agggtcagcg 300 tatgaccatc aggttggcaa atcaggcacggctaacatgg ttgcaaccat gctcacccaa 360 ggaactgaca gcctttctga agatgagtttgttgctgcca aagagcgtct tggcattgat 420 tttaccagta cagcaaataa ggataacttaactttatcat taagaagctt gtctgatcaa 480 tcattattaa atcaagccgc cgatttaatggtcgatgctg tcactcaacc tgcttttgat 540 gataagactc tacaacgcaa caaaaatcagctcatcacca gtttaaaaca aaaaaagcaa 600 aacccttatc atgtagcttc tgttgcttatcatcaagccg tatatgaaaa tcatccttat 660 gcacacgcaa ccacaggcga tgaagatagtattgccaaaa ttgatcgtga tgagctgctt 720 aatttttggc atacttttat taatgcaaataatgcgacac tggtgattac aggtgatatg 780 accgccgagc aagccaaatc acttgccaaccatctgaccg ccaaattacc gacaggcaag 840 tcgtataaaa atacgctgga tttgacaaaaccagttaagg ctcgtcatat ccatattcct 900 cacaacagta gtcaaaccca aatcatcatcggtcatccca ccagtaaagt acgcacggac 960 aaagcaggtc gtcaagagtt cagcgatttttcattaggta atgaaatttt ggcaggtggt 1020 gattttaatg ccagattgat gaaaaccattcgagagcaaa aaggctacac ttatggcatt 1080 tatggcggta tggaacgcct cagagcaggtggtaattatg tggttgaatt ttcaaccgat 1140 ggcgataaag cagccgatgc cattttagagacgctacaca tcattaatga gtcgctgaat 1200 gaaggcataa cccaagaaga gcttgagttggtgcgtttgg gcaataaaaa tggttttgcc 1260 aatatttttt caagcaatgc cagtattcatcgtgtcattg gtgctttatt tgttgccgat 1320 tatccaaaag atcatcttaa ccatacgctcaatcgcttgg ataatgccac gataaatagt 1380 gttaataccg cactgaactt gcgtatcaagcctgatgaat ttatcatcat caccgtgggt 1440 aaaactaagc caaatttgga caaataa 14672 488 PRT Moraxella catarrhalis 2 Met Asp Thr Asp Met Lys His Leu ThrLys His Arg Leu Ser Ala Ala 1 5 10 15 Ile Ile Gly Val Leu Leu Phe IleSer Pro Ser Val Gln Ala Asn Thr 20 25 30 Thr His His His Thr Leu Thr SerSer Glu Leu Lys Leu Ala Asp Asp 35 40 45 Ser Ile Ile Asp Ser Ile Asn GlnLeu Gly Glu Leu Thr Val Asn Ile 50 55 60 Pro Asn Thr Gln Tyr Phe Gln ThrAsn Asn Gly Val Ser Val Ala Phe 65 70 75 80 Thr Pro Leu His Glu Leu ProIle Val Asp Ile Ser Leu Tyr Phe Asn 85 90 95 Ala Gly Ser Ala Tyr Asp HisGln Val Gly Lys Ser Gly Thr Ala Asn 100 105 110 Met Val Ala Thr Met LeuThr Gln Gly Thr Asp Ser Leu Ser Glu Asp 115 120 125 Glu Phe Val Ala AlaLys Glu Arg Leu Gly Ile Asp Phe Thr Ser Thr 130 135 140 Ala Asn Lys AspAsn Leu Thr Leu Ser Leu Arg Ser Leu Ser Asp Gln 145 150 155 160 Ser LeuLeu Asn Gln Ala Ala Asp Leu Met Val Asp Ala Val Thr Gln 165 170 175 ProAla Phe Asp Asp Lys Thr Leu Gln Arg Asn Lys Asn Gln Leu Ile 180 185 190Thr Ser Leu Lys Gln Lys Lys Gln Asn Pro Tyr His Val Ala Ser Val 195 200205 Ala Tyr His Gln Ala Val Tyr Glu Asn His Pro Tyr Ala His Ala Thr 210215 220 Thr Gly Asp Glu Asp Ser Ile Ala Lys Ile Asp Arg Asp Glu Leu Leu225 230 235 240 Asn Phe Trp His Thr Phe Ile Asn Ala Asn Asn Ala Thr LeuVal Ile 245 250 255 Thr Gly Asp Met Thr Ala Glu Gln Ala Lys Ser Leu AlaAsn His Leu 260 265 270 Thr Ala Lys Leu Pro Thr Gly Lys Ser Tyr Lys AsnThr Leu Asp Leu 275 280 285 Thr Lys Pro Val Lys Ala Arg His Ile His IlePro His Asn Ser Ser 290 295 300 Gln Thr Gln Ile Ile Ile Gly His Pro ThrSer Lys Val Arg Thr Asp 305 310 315 320 Lys Ala Gly Arg Gln Glu Phe SerAsp Phe Ser Leu Gly Asn Glu Ile 325 330 335 Leu Ala Gly Gly Asp Phe AsnAla Arg Leu Met Lys Thr Ile Arg Glu 340 345 350 Gln Lys Gly Tyr Thr TyrGly Ile Tyr Gly Gly Met Glu Arg Leu Arg 355 360 365 Ala Gly Gly Asn TyrVal Val Glu Phe Ser Thr Asp Gly Asp Lys Ala 370 375 380 Ala Asp Ala IleLeu Glu Thr Leu His Ile Ile Asn Glu Ser Leu Asn 385 390 395 400 Glu GlyIle Thr Gln Glu Glu Leu Glu Leu Val Arg Leu Gly Asn Lys 405 410 415 AsnGly Phe Ala Asn Ile Phe Ser Ser Asn Ala Ser Ile His Arg Val 420 425 430Ile Gly Ala Leu Phe Val Ala Asp Tyr Pro Lys Asp His Leu Asn His 435 440445 Thr Leu Asn Arg Leu Asp Asn Ala Thr Ile Asn Ser Val Asn Thr Ala 450455 460 Leu Asn Leu Arg Ile Lys Pro Asp Glu Phe Ile Ile Ile Thr Val Gly465 470 475 480 Lys Thr Lys Pro Asn Leu Asp Lys 485 3 1299 DNA Moraxellacatarrhalis 3 gagcttaaac ttgctgatga tagtattatt gatagtatca atcaattgggtgagctgacc 60 gtcaatattc caaatacaca atattttcaa accaacaacg gtgtgagcgttgcttttacg 120 ccattacatg agctgcctat tgtcgatatc agcttgtatt ttaatgcagggtcagcgtat 180 gaccatcagg ttggcaaatc aggcacggct aacatggttg caaccatgctcacccaagga 240 actgacagcc tttctgaaga tgagtttgtt gctgccaaag agcgtcttggcattgatttt 300 accagtacag caaataagga taacttaact ttatcattaa gaagcttgtctgatcaatca 360 ttattaaatc aagccgccga tttaatggtc gatgctgtca ctcaacctgcttttgatgat 420 aagactctac aacgcaacaa aaatcagctc atcaccagtt taaaacaaaaaaagcaaaac 480 ccttatcatg tagcttctgt tgcttatcat caagccgtat atgaaaatcatccttatgca 540 cacgcaacca caggcgatga agatagtatt gccaaaattg atcgtgatgagctgcttaat 600 ttttggcata cttttattaa tgcaaataat gcgacactgg tgattacaggtgatatgacc 660 gccgagcaag ccaaatcact tgccaaccat ctgaccgcca aattaccgacaggcaagtcg 720 tataaaaata cgctggattt gacaaaacca gttaaggctc gccatatccatattcctcac 780 aacagtagtc aaacccaaat catcatcggt caccccacca gtaaagtacgcacggacaaa 840 gcaggtcgtc aagagttcag cgatttttca ttaggtaatg aaattttggcaggtggtgat 900 tttaatgcca gattgatgaa aaccattcga gagcaaaaag gctacacttatggcatttat 960 ggcggtatgg aacgcctcag agcaggtggt aattatgtgg ttgaattttcaaccgatggc 1020 gataaagcag ccgatgccat tttagagacg ctacacatca ttaatgagtcgctgaatgaa 1080 ggcataaccc aagaagagct tgaattggtg cgtttgggta ataaaaatggttttgccaat 1140 attttttcaa gcaatgccag tattcatcgt gtcattggtg ctttatttgttgccgattat 1200 ccaaaagatc atcttaacca tacgctcaat cgcttggata atgccacgataaatagtgtt 1260 aataccgcac tgaacttgcg tatcaagcct gatgaattt 1299 4 433PRT Moraxella catarrhalis 4 Glu Leu Lys Leu Ala Asp Asp Ser Ile Ile AspSer Ile Asn Gln Leu 1 5 10 15 Gly Glu Leu Thr Val Asn Ile Pro Asn ThrGln Tyr Phe Gln Thr Asn 20 25 30 Asn Gly Val Ser Val Ala Phe Thr Pro LeuHis Glu Leu Pro Ile Val 35 40 45 Asp Ile Ser Leu Tyr Phe Asn Ala Gly SerAla Tyr Asp His Gln Val 50 55 60 Gly Lys Ser Gly Thr Ala Asn Met Val AlaThr Met Leu Thr Gln Gly 65 70 75 80 Thr Asp Ser Leu Ser Glu Asp Glu PheVal Ala Ala Lys Glu Arg Leu 85 90 95 Gly Ile Asp Phe Thr Ser Thr Ala AsnLys Asp Asn Leu Thr Leu Ser 100 105 110 Leu Arg Ser Leu Ser Asp Gln SerLeu Leu Asn Gln Ala Ala Asp Leu 115 120 125 Met Val Asp Ala Val Thr GlnPro Ala Phe Asp Asp Lys Thr Leu Gln 130 135 140 Arg Asn Lys Asn Gln LeuIle Thr Ser Leu Lys Gln Lys Lys Gln Asn 145 150 155 160 Pro Tyr His ValAla Ser Val Ala Tyr His Gln Ala Val Tyr Glu Asn 165 170 175 His Pro TyrAla His Ala Thr Thr Gly Asp Glu Asp Ser Ile Ala Lys 180 185 190 Ile AspArg Asp Glu Leu Leu Asn Phe Trp His Thr Phe Ile Asn Ala 195 200 205 AsnAsn Ala Thr Leu Val Ile Thr Gly Asp Met Thr Ala Glu Gln Ala 210 215 220Lys Ser Leu Ala Asn His Leu Thr Ala Lys Leu Pro Thr Gly Lys Ser 225 230235 240 Tyr Lys Asn Thr Leu Asp Leu Thr Lys Pro Val Lys Ala Arg His Ile245 250 255 His Ile Pro His Asn Ser Ser Gln Thr Gln Ile Ile Ile Gly HisPro 260 265 270 Thr Ser Lys Val Arg Thr Asp Lys Ala Gly Arg Gln Glu PheSer Asp 275 280 285 Phe Ser Leu Gly Asn Glu Ile Leu Ala Gly Gly Asp PheAsn Ala Arg 290 295 300 Leu Met Lys Thr Ile Arg Glu Gln Lys Gly Tyr ThrTyr Gly Ile Tyr 305 310 315 320 Gly Gly Met Glu Arg Leu Arg Ala Gly GlyAsn Tyr Val Val Glu Phe 325 330 335 Ser Thr Asp Gly Asp Lys Ala Ala AspAla Ile Leu Glu Thr Leu His 340 345 350 Ile Ile Asn Glu Ser Leu Asn GluGly Ile Thr Gln Glu Glu Leu Glu 355 360 365 Leu Val Arg Leu Gly Asn LysAsn Gly Phe Ala Asn Ile Phe Ser Ser 370 375 380 Asn Ala Ser Ile His ArgVal Ile Gly Ala Leu Phe Val Ala Asp Tyr 385 390 395 400 Pro Lys Asp HisLeu Asn His Thr Leu Asn Arg Leu Asp Asn Ala Thr 405 410 415 Ile Asn SerVal Asn Thr Ala Leu Asn Leu Arg Ile Lys Pro Asp Glu 420 425 430 Phe 51299 DNA Moraxella catarrhalis 5 gagcttaaac ttgctgatga tagtattattgatagtatca atcaattggg tgagctgacc 60 gtcaatattc caaatacaca atattttcaaaccaacaacg gtgtgagcgt tgcttttacg 120 ccattacatg agctgcctat tgtcgatatcagcttgtatt ttaatgcagg gtcagcgtat 180 gaccatcagg ttggcaaatc aggcacggctaacatggttg caaccatgct cacccaagga 240 actgacagcc tttctgaaga tgagtttgttgctgccaaag agcgtcttgg cattgatttt 300 accagtacag caaataagga taacttaactttatcattaa gaagcttgtc tgatcaatca 360 ttattaaatc aagccgccga tttaatggtcgatgctgtca ctcaacctgc ttttgatgat 420 aagactctac aacgcaacaa aaatcagctcatcaccagtt taaaacaaaa aaagcaaaac 480 ccttatcatg tagcttctgt tgcttatcatcaagccgtat atgaaaatca tccttatgca 540 cacgcaacca caggcgatga agatagtattgccaaaattg atcgtgatga gctgcttaat 600 ttttggcata cttttattaa tgcaaataatgcgacactgg tgattacagg tgatatgacc 660 gccgagcaag ccaaatcact tgccaaccatctgaccgcca aattaccgac aggcaagtct 720 tataaaaata cgctggattt gacaaaaccagttaaggctc gccatatcca tattcctcac 780 aacagtagtc aaacccaaat catcatcggtcaccccacca gtaaagtacg cacggacaaa 840 gcaggtcgtc aagagttcag cgatttttcattaggtaatg aaattttggc aggtggtgat 900 tttaatgcca gattgatgaa aaccattcgagagcaaaaag gctacactta tggcatttat 960 ggcggtatgg aacgcctcag agcaggtggtaattatgtgg ttgaattttc aaccgatggc 1020 gataaagcag ccgatgccat tttagagacgctacacatca ttaatgagtc gctgaatgaa 1080 ggcataaccc aagaagagct tgagttggtgcgtttgggca ataaaaatgg ttttgccaat 1140 attttttcaa gcaatgccag tattcatcgtgtcattggtg ctttatttgt tgccgattat 1200 ccaaaagatc atcttaacca tacgctcaatcgcttggata atgccacgat aaatagtgtt 1260 aataccgcac tgaacttgcg tatcaagcctgatgaattt 1299 6 433 PRT Moraxella catarrhalis 6 Glu Leu Lys Leu Ala AspAsp Ser Ile Ile Asp Ser Ile Asn Gln Leu 1 5 10 15 Gly Glu Leu Thr ValAsn Ile Pro Asn Thr Gln Tyr Phe Gln Thr Asn 20 25 30 Asn Gly Val Ser ValAla Phe Thr Pro Leu His Glu Leu Pro Ile Val 35 40 45 Asp Ile Ser Leu TyrPhe Asn Ala Gly Ser Ala Tyr Asp His Gln Val 50 55 60 Gly Lys Ser Gly ThrAla Asn Met Val Ala Thr Met Leu Thr Gln Gly 65 70 75 80 Thr Asp Ser LeuSer Glu Asp Glu Phe Val Ala Ala Lys Glu Arg Leu 85 90 95 Gly Ile Asp PheThr Ser Thr Ala Asn Lys Asp Asn Leu Thr Leu Ser 100 105 110 Leu Arg SerLeu Ser Asp Gln Ser Leu Leu Asn Gln Ala Ala Asp Leu 115 120 125 Met ValAsp Ala Val Thr Gln Pro Ala Phe Asp Asp Lys Thr Leu Gln 130 135 140 ArgAsn Lys Asn Gln Leu Ile Thr Ser Leu Lys Gln Lys Lys Gln Asn 145 150 155160 Pro Tyr His Val Ala Ser Val Ala Tyr His Gln Ala Val Tyr Glu Asn 165170 175 His Pro Tyr Ala His Ala Thr Thr Gly Asp Glu Asp Ser Ile Ala Lys180 185 190 Ile Asp Arg Asp Glu Leu Leu Asn Phe Trp His Thr Phe Ile AsnAla 195 200 205 Asn Asn Ala Thr Leu Val Ile Thr Gly Asp Met Thr Ala GluGln Ala 210 215 220 Lys Ser Leu Ala Asn His Leu Thr Ala Lys Leu Pro ThrGly Lys Ser 225 230 235 240 Tyr Lys Asn Thr Leu Asp Leu Thr Lys Pro ValLys Ala Arg His Ile 245 250 255 His Ile Pro His Asn Ser Ser Gln Thr GlnIle Ile Ile Gly His Pro 260 265 270 Thr Ser Lys Val Arg Thr Asp Lys AlaGly Arg Gln Glu Phe Ser Asp 275 280 285 Phe Ser Leu Gly Asn Glu Ile LeuAla Gly Gly Asp Phe Asn Ala Arg 290 295 300 Leu Met Lys Thr Ile Arg GluGln Lys Gly Tyr Thr Tyr Gly Ile Tyr 305 310 315 320 Gly Gly Met Glu ArgLeu Arg Ala Gly Gly Asn Tyr Val Val Glu Phe 325 330 335 Ser Thr Asp GlyAsp Lys Ala Ala Asp Ala Ile Leu Glu Thr Leu His 340 345 350 Ile Ile AsnGlu Ser Leu Asn Glu Gly Ile Thr Gln Glu Glu Leu Glu 355 360 365 Leu ValArg Leu Gly Asn Lys Asn Gly Phe Ala Asn Ile Phe Ser Ser 370 375 380 AsnAla Ser Ile His Arg Val Ile Gly Ala Leu Phe Val Ala Asp Tyr 385 390 395400 Pro Lys Asp His Leu Asn His Thr Leu Asn Arg Leu Asp Asn Ala Thr 405410 415 Ile Asn Ser Val Asn Thr Ala Leu Asn Leu Arg Ile Lys Pro Asp Glu420 425 430 Phe 7 1299 DNA Moraxella catarrhalis 7 gagcttaaac ttgctgatgatagtattatt gatagtatca atcaattggg tgagctgacc 60 gtcaatattc caaatacacaatattttcaa accaacaacg gtgtgagcgt tgcttttacg 120 ccattacatg agctgcctattgtcgatatc agcttgtatt ttaatgcagg gtcagcgtat 180 gaccatcagg ttggcaaatcaggcacggct aacatggttg caaccatgct cacccaagga 240 actgacagcc tttctgaagatgagtttgtt gctgccaaag agcgtcttgg cattgatttt 300 accagtacag caaataaggataacttaact ttatcattaa gaagcttgtc tgatcaatca 360 ttattaaatc aagccgccgatttaatggtc gatgctgtca ctcaacctgc ttttgatgat 420 aagactctac aacgcaacaaaaatcagctc atcaccagtt taaaacaaaa aaagcaaaac 480 ccttatcatg tagcttctgttgcttatcat caagccgtat atgaaaatca tccttatgca 540 cacgcaacca caggcgatgaagatagtatt gccaaaattg atcgtgatga gctgcttaat 600 ttttggcata cttttattaatgcaaataat gcgacactgg tgattacagg tgatatgacc 660 gccgagcaag ccaaatcacttgccaaccat ctgaccgcca aattaccgac aggcaagtcg 720 tataaaaata cgctggatttgacaaaacca gttaaggctc gccatatcca tattcctcac 780 aacagtagtc aaacccaaatcatcatcggt caccccacca gtaaagtacg cacggacaaa 840 gcaggtcgtc aagagttcagcgatttttca ttaggtaatg aaattttggc aggtggtgat 900 tttaatgcca gattgatgaaaaccattcga gagcaaaaag gctacactta tggcatttat 960 ggcggtatgg aacgcctcagagcaggtggt aattatgtgg ttgaattttc aaccgatggc 1020 gataaagcag ccgatgccattttagagacg ctacacatca ttaatgagtc gctgaatgaa 1080 ggcataaccc aagaagagcttgaattggtg cgtttgggta ataaaaatgg ttttgccaat 1140 attttttcaa gcaatgccagtattcatcgt gtcattggtg ctttatttgt tgccgattat 1200 ccaaaagacc atcttaaccatacgctcaat cgcttggata atgccacgat aaatagtgtt 1260 aataccgcac tgaacttgcgtatcaagcct gatgaattt 1299 8 433 PRT Moraxella catarrhalis 8 Glu Leu LysLeu Ala Asp Asp Ser Ile Ile Asp Ser Ile Asn Gln Leu 1 5 10 15 Gly GluLeu Thr Val Asn Ile Pro Asn Thr Gln Tyr Phe Gln Thr Asn 20 25 30 Asn GlyVal Ser Val Ala Phe Thr Pro Leu His Glu Leu Pro Ile Val 35 40 45 Asp IleSer Leu Tyr Phe Asn Ala Gly Ser Ala Tyr Asp His Gln Val 50 55 60 Gly LysSer Gly Thr Ala Asn Met Val Ala Thr Met Leu Thr Gln Gly 65 70 75 80 ThrAsp Ser Leu Ser Glu Asp Glu Phe Val Ala Ala Lys Glu Arg Leu 85 90 95 GlyIle Asp Phe Thr Ser Thr Ala Asn Lys Asp Asn Leu Thr Leu Ser 100 105 110Leu Arg Ser Leu Ser Asp Gln Ser Leu Leu Asn Gln Ala Ala Asp Leu 115 120125 Met Val Asp Ala Val Thr Gln Pro Ala Phe Asp Asp Lys Thr Leu Gln 130135 140 Arg Asn Lys Asn Gln Leu Ile Thr Ser Leu Lys Gln Lys Lys Gln Asn145 150 155 160 Pro Tyr His Val Ala Ser Val Ala Tyr His Gln Ala Val TyrGlu Asn 165 170 175 His Pro Tyr Ala His Ala Thr Thr Gly Asp Glu Asp SerIle Ala Lys 180 185 190 Ile Asp Arg Asp Glu Leu Leu Asn Phe Trp His ThrPhe Ile Asn Ala 195 200 205 Asn Asn Ala Thr Leu Val Ile Thr Gly Asp MetThr Ala Glu Gln Ala 210 215 220 Lys Ser Leu Ala Asn His Leu Thr Ala LysLeu Pro Thr Gly Lys Ser 225 230 235 240 Tyr Lys Asn Thr Leu Asp Leu ThrLys Pro Val Lys Ala Arg His Ile 245 250 255 His Ile Pro His Asn Ser SerGln Thr Gln Ile Ile Ile Gly His Pro 260 265 270 Thr Ser Lys Val Arg ThrAsp Lys Ala Gly Arg Gln Glu Phe Ser Asp 275 280 285 Phe Ser Leu Gly AsnGlu Ile Leu Ala Gly Gly Asp Phe Asn Ala Arg 290 295 300 Leu Met Lys ThrIle Arg Glu Gln Lys Gly Tyr Thr Tyr Gly Ile Tyr 305 310 315 320 Gly GlyMet Glu Arg Leu Arg Ala Gly Gly Asn Tyr Val Val Glu Phe 325 330 335 SerThr Asp Gly Asp Lys Ala Ala Asp Ala Ile Leu Glu Thr Leu His 340 345 350Ile Ile Asn Glu Ser Leu Asn Glu Gly Ile Thr Gln Glu Glu Leu Glu 355 360365 Leu Val Arg Leu Gly Asn Lys Asn Gly Phe Ala Asn Ile Phe Ser Ser 370375 380 Asn Ala Ser Ile His Arg Val Ile Gly Ala Leu Phe Val Ala Asp Tyr385 390 395 400 Pro Lys Asp His Leu Asn His Thr Leu Asn Arg Leu Asp AsnAla Thr 405 410 415 Ile Asn Ser Val Asn Thr Ala Leu Asn Leu Arg Ile LysPro Asp Glu 420 425 430 Phe 9 1656 DNA Moraxella catarrhalis 9atgagcttaa ttaataaatt aaatgaacgc attacgccgc atgtcttaac ttcgattaaa 60aatcaagatg gcgataatgc tgataaatct aatttgttaa ccgcatttta taccattttt 120gcaggacgct tgagtaatga agatgtgtat cagcgtgcca atgctttgcc tgataatgag 180cttgagcatg ggcatcatct gctcaatgtt gcttttagtg atgtttcaac tggtgaagat 240cagattgctt ctttgagtaa tcaattagcc gatgaatatc atgtttcgcc agtaacggca 300cgcaccgcaa tcgcaacggc agcacctttg gctttggcac gcattaaaga gcaagcaggt 360gcattatctg taccgtcttt tattcgtact caattggcta aagaagaaaa ccgtttgcca 420acttgggcgc atactttatt gccagcaggg ctatttgcaa ccgctgccac aaccaccgcc 480gagcctgtaa cgacagcctc tgctgttgtg aaagagcctg tcaaaccaag tgttgtgaca 540gaaccagttc atccagctgc ggctaccacc ccagtcaaaa caccaactgc ccagcattac 600gaaaacaaag aaaaaagtcc ttttctaaaa acgattctac cgattattgg attgattatt 660tttgcaggct tggcatggct tttgttaaga gcatgtcaag acaaaccaac acctgttgcg 720gcacctgttg cgacagatac agcacctgtg gtagcggata atgctgtaca ggcagaccca 780acacaaacag gtgttgccca agcacctgca acgcttagct tgtctgttga tgaaacgggt 840caagcgttgt actcgcaccg tgctcaggtt ggtagtgaag agcttgcagg tcatatccgt 900gcagctattg ctcaagtctt tggcgtacaa gatttaacca ttcaaaatac caatgtacat 960accgctacga tgccagcggc agaatactta ccagcaattt tgggtttgat gaaaggtgta 1020ccaaattcaa gcgttgtgat tcatgatcat acggtacgct ttaatgcaac cacgccagaa 1080gatgtagcaa aactggtaga gggtgctaaa aatattctac ccgctgattt tactgtagaa 1140gcagaacctg aacttgatat taatactgcg gttgccgata gtattgaaac agcgcgtgtt 1200gctattgttg ctttgggtga tacggttgaa gaaaatgaga tggatatttt aatcaatgca 1260ttaaataccc aaatcattaa ctttgcttta gactcaaccg aaattcccca agaaaataaa 1320gaaatcttgg atttggctgc cgaaaaatta aaggcagtgc ctgaaacaac tttgcgtatc 1380attggtcata cagacactca aggcacacat gagtataatc aagatttatc agaatctcgt 1440gctgctgctg ttaaagagta tttggtatca aaaggtgttg ctgctgaacg cttgaacact 1500caaggtgcaa gttttgatta tccagttgca tcaaatgcta ccgaacaagg tcgcttccaa 1560aaccgtcgta ttgagtttgt acttttccaa gaaggtgaag caattactca agtcggtcat 1620gctgaagatg caccaacacc tgttgcacaa aactga 1656 10 551 PRT Moraxellacatarrhalis 10 Met Ser Leu Ile Asn Lys Leu Asn Glu Arg Ile Thr Pro HisVal Leu 1 5 10 15 Thr Ser Ile Lys Asn Gln Asp Gly Asp Asn Ala Asp LysSer Asn Leu 20 25 30 Leu Thr Ala Phe Tyr Thr Ile Phe Ala Gly Arg Leu SerAsn Glu Asp 35 40 45 Val Tyr Gln Arg Ala Asn Ala Leu Pro Asp Asn Glu LeuGlu His Gly 50 55 60 His His Leu Leu Asn Val Ala Phe Ser Asp Val Ser ThrGly Glu Asp 65 70 75 80 Gln Ile Ala Ser Leu Ser Asn Gln Leu Ala Asp GluTyr His Val Ser 85 90 95 Pro Val Thr Ala Arg Thr Ala Ile Ala Thr Ala AlaPro Leu Ala Leu 100 105 110 Ala Arg Ile Lys Glu Gln Ala Gly Ala Leu SerVal Pro Ser Phe Ile 115 120 125 Arg Thr Gln Leu Ala Lys Glu Glu Asn ArgLeu Pro Thr Trp Ala His 130 135 140 Thr Leu Leu Pro Ala Gly Leu Phe AlaThr Ala Ala Thr Thr Thr Ala 145 150 155 160 Glu Pro Val Thr Thr Ala SerAla Val Val Lys Glu Pro Val Lys Pro 165 170 175 Ser Val Val Thr Glu ProVal His Pro Ala Ala Ala Thr Thr Pro Val 180 185 190 Lys Thr Pro Thr AlaGln His Tyr Glu Asn Lys Glu Lys Ser Pro Phe 195 200 205 Leu Lys Thr IleLeu Pro Ile Ile Gly Leu Ile Ile Phe Ala Gly Leu 210 215 220 Ala Trp LeuLeu Leu Arg Ala Cys Gln Asp Lys Pro Thr Pro Val Ala 225 230 235 240 AlaPro Val Ala Thr Asp Thr Ala Pro Val Val Ala Asp Asn Ala Val 245 250 255Gln Ala Asp Pro Thr Gln Thr Gly Val Ala Gln Ala Pro Ala Thr Leu 260 265270 Ser Leu Ser Val Asp Glu Thr Gly Gln Ala Leu Tyr Ser His Arg Ala 275280 285 Gln Val Gly Ser Glu Glu Leu Ala Gly His Ile Arg Ala Ala Ile Ala290 295 300 Gln Val Phe Gly Val Gln Asp Leu Thr Ile Gln Asn Thr Asn ValHis 305 310 315 320 Thr Ala Thr Met Pro Ala Ala Glu Tyr Leu Pro Ala IleLeu Gly Leu 325 330 335 Met Lys Gly Val Pro Asn Ser Ser Val Val Ile HisAsp His Thr Val 340 345 350 Arg Phe Asn Ala Thr Thr Pro Glu Asp Val AlaLys Leu Val Glu Gly 355 360 365 Ala Lys Asn Ile Leu Pro Ala Asp Phe ThrVal Glu Ala Glu Pro Glu 370 375 380 Leu Asp Ile Asn Thr Ala Val Ala AspSer Ile Glu Thr Ala Arg Val 385 390 395 400 Ala Ile Val Ala Leu Gly AspThr Val Glu Glu Asn Glu Met Asp Ile 405 410 415 Leu Ile Asn Ala Leu AsnThr Gln Ile Ile Asn Phe Ala Leu Asp Ser 420 425 430 Thr Glu Ile Pro GlnGlu Asn Lys Glu Ile Leu Asp Leu Ala Ala Glu 435 440 445 Lys Leu Lys AlaVal Pro Glu Thr Thr Leu Arg Ile Ile Gly His Thr 450 455 460 Asp Thr GlnGly Thr His Glu Tyr Asn Gln Asp Leu Ser Glu Ser Arg 465 470 475 480 AlaAla Ala Val Lys Glu Tyr Leu Val Ser Lys Gly Val Ala Ala Glu 485 490 495Arg Leu Asn Thr Gln Gly Ala Ser Phe Asp Tyr Pro Val Ala Ser Asn 500 505510 Ala Thr Glu Gln Gly Arg Phe Gln Asn Arg Arg Ile Glu Phe Val Leu 515520 525 Phe Gln Glu Gly Glu Ala Ile Thr Gln Val Gly His Ala Glu Asp Ala530 535 540 Pro Thr Pro Val Ala Gln Asn 545 550 11 1251 DNA Moraxellacatarrhalis 11 atggatacaa aacacattca gcaaaattgg cttctacctg atggtgtggctgatgtacta 60 tttaccgatg ctcaaaaaca agaaagcctg cgtgatgcct tgctatttgtgctaaccgca 120 cacggttatc gcttggtgtc accaccatta atagagtata ccgaaagtctgctaaataat 180 gctgacgaag atctaaaacg ccaaactttc aaatttatcg atcagctcaatggtcgtttg 240 atgggtttgc gtgccgatat tacgccacaa attctacgca ttgatagcaaatatggtcaa 300 ggcatcagcc gttactgtta tgttgggcaa gttgtcaaaa ccctaccgactggtctgttt 360 gggctgcgta caccgcttca attgggtgct gagatttttg ggatagatgatatccgtgcc 420 gagcttgagc tgattgatct attggccgca ttggcagatg agatcggactaggccgagag 480 atgctacatg tggatattgg tcatgtcgct atttttgatc gcttgtgtcagttgcatggc 540 gtttcaaata aagatgctga tgagctgatt ggcatttacc ataaaaaagccatgccagaa 600 cttgccaaat ggtgccaaaa tattggcaat agcctaaaca gcccaagcgatgcaaccgat 660 tttttggtat tggctaagca tacattaagc agtgatcgga caccaaatgccgaggcttta 720 ttaagtaaac tgtccgataa agctcgccaa gataataaaa tcatccaagcggcaaatgag 780 cttgctactt tggcggcaca tatcagagcg gtgggtatga gtgtgagtattgatgtgact 840 gaattgtcag gatatcatta tcatactggt gtggtattta atgtctatttgggtaataga 900 accacacaga ctcaagcttt ggtacgaggc ggtcgctttg atggtatctcaactcacagc 960 gtagcaaggg gcgcaactgg ttttagcatg gatattaatc gtttgcttgaatttgtagag 1020 cttgaagaag atactgtgat tttggtggat tatcacgatt tgcaaaatgctgatgcagac 1080 acaaaagctg atttggccac acaaattaaa accttgcaat ctgaaggctgtattgtcatt 1140 aagcctttga ctgtagatga taagcctaac cagattgatg gtgttttgcattgggacacc 1200 gatcaagata agccgatttg ggcggtgcga ttagttggtg atgagtacta a1251 12 416 PRT Moraxella catarrhalis 12 Met Asp Thr Lys His Ile Gln GlnAsn Trp Leu Leu Pro Asp Gly Val 1 5 10 15 Ala Asp Val Leu Phe Thr AspAla Gln Lys Gln Glu Ser Leu Arg Asp 20 25 30 Ala Leu Leu Phe Val Leu ThrAla His Gly Tyr Arg Leu Val Ser Pro 35 40 45 Pro Leu Ile Glu Tyr Thr GluSer Leu Leu Asn Asn Ala Asp Glu Asp 50 55 60 Leu Lys Arg Gln Thr Phe LysPhe Ile Asp Gln Leu Asn Gly Arg Leu 65 70 75 80 Met Gly Leu Arg Ala AspIle Thr Pro Gln Ile Leu Arg Ile Asp Ser 85 90 95 Lys Tyr Gly Gln Gly IleSer Arg Tyr Cys Tyr Val Gly Gln Val Val 100 105 110 Lys Thr Leu Pro ThrGly Leu Phe Gly Leu Arg Thr Pro Leu Gln Leu 115 120 125 Gly Ala Glu IlePhe Gly Ile Asp Asp Ile Arg Ala Glu Leu Glu Leu 130 135 140 Ile Asp LeuLeu Ala Ala Leu Ala Asp Glu Ile Gly Leu Gly Arg Glu 145 150 155 160 MetLeu His Val Asp Ile Gly His Val Ala Ile Phe Asp Arg Leu Cys 165 170 175Gln Leu His Gly Val Ser Asn Lys Asp Ala Asp Glu Leu Ile Gly Ile 180 185190 Tyr His Lys Lys Ala Met Pro Glu Leu Ala Lys Trp Cys Gln Asn Ile 195200 205 Gly Asn Ser Leu Asn Ser Pro Ser Asp Ala Thr Asp Phe Leu Val Leu210 215 220 Ala Lys His Thr Leu Ser Ser Asp Arg Thr Pro Asn Ala Glu AlaLeu 225 230 235 240 Leu Ser Lys Leu Ser Asp Lys Ala Arg Gln Asp Asn LysIle Ile Gln 245 250 255 Ala Ala Asn Glu Leu Ala Thr Leu Ala Ala His IleArg Ala Val Gly 260 265 270 Met Ser Val Ser Ile Asp Val Thr Glu Leu SerGly Tyr His Tyr His 275 280 285 Thr Gly Val Val Phe Asn Val Tyr Leu GlyAsn Arg Thr Thr Gln Thr 290 295 300 Gln Ala Leu Val Arg Gly Gly Arg PheAsp Gly Ile Ser Thr His Ser 305 310 315 320 Val Ala Arg Gly Ala Thr GlyPhe Ser Met Asp Ile Asn Arg Leu Leu 325 330 335 Glu Phe Val Glu Leu GluGlu Asp Thr Val Ile Leu Val Asp Tyr His 340 345 350 Asp Leu Gln Asn AlaAsp Ala Asp Thr Lys Ala Asp Leu Ala Thr Gln 355 360 365 Ile Lys Thr LeuGln Ser Glu Gly Cys Ile Val Ile Lys Pro Leu Thr 370 375 380 Val Asp AspLys Pro Asn Gln Ile Asp Gly Val Leu His Trp Asp Thr 385 390 395 400 AspGln Asp Lys Pro Ile Trp Ala Val Arg Leu Val Gly Asp Glu Tyr 405 410 41513 639 DNA Moraxella catarrhalis 13 atgaataatt ttgtgtatca gctacaaagtttttggtatg agcttaatca ggtcaatcgt 60 cataccattg ctcaatcacc caaatatatacagctgacgg tacttggttt gatcgtgatg 120 atcattggca tttttggctg gctacttgcgattttaccaa ccattcaaaa gcttaatgca 180 gcccaaagtc aagaatctgc cttaattgatgaatttgcca ctaaatatca taaagcccag 240 cagtttgacc atctaagcca tcaggtcatacaaaaaaata cacaacttga aaatcagctc 300 aatgctctgc cacgcacagc accgatgagcgagattatcg gaatgataaa taccaaagca 360 caagcggtta atgtgcaggt ggtgagtgcatcagttcaag caggtcgtga acaggattat 420 tataccgaac gccctatcgc agtgagtgcgacaggggatt atcatgcttt gggtcgatgg 480 ttacttgagt tgtcagaggc taaccatttgctgacagtgc atgattttga tctgaaggct 540 ggtttgaacc atcagctgat gatgattgctcagatgaaaa cttatcaagc aaacaaacgc 600 ccaaaaccag ttgctcagca ggtgcctgatgttcaatga 639 14 212 PRT Moraxella catarrhalis 14 Met Asn Asn Phe ValTyr Gln Leu Gln Ser Phe Trp Tyr Glu Leu Asn 1 5 10 15 Gln Val Asn ArgHis Thr Ile Ala Gln Ser Pro Lys Tyr Ile Gln Leu 20 25 30 Thr Val Leu GlyLeu Ile Val Met Ile Ile Gly Ile Phe Gly Trp Leu 35 40 45 Leu Ala Ile LeuPro Thr Ile Gln Lys Leu Asn Ala Ala Gln Ser Gln 50 55 60 Glu Ser Ala LeuIle Asp Glu Phe Ala Thr Lys Tyr His Lys Ala Gln 65 70 75 80 Gln Phe AspHis Leu Ser His Gln Val Ile Gln Lys Asn Thr Gln Leu 85 90 95 Glu Asn GlnLeu Asn Ala Leu Pro Arg Thr Ala Pro Met Ser Glu Ile 100 105 110 Ile GlyMet Ile Asn Thr Lys Ala Gln Ala Val Asn Val Gln Val Val 115 120 125 SerAla Ser Val Gln Ala Gly Arg Glu Gln Asp Tyr Tyr Thr Glu Arg 130 135 140Pro Ile Ala Val Ser Ala Thr Gly Asp Tyr His Ala Leu Gly Arg Trp 145 150155 160 Leu Leu Glu Leu Ser Glu Ala Asn His Leu Leu Thr Val His Asp Phe165 170 175 Asp Leu Lys Ala Gly Leu Asn His Gln Leu Met Met Ile Ala GlnMet 180 185 190 Lys Thr Tyr Gln Ala Asn Lys Arg Pro Lys Pro Val Ala GlnGln Val 195 200 205 Pro Asp Val Gln 210 15 35 DNA Artificial primer 15catcagtgca tatgaatacg acacaccatc acacg 35 16 40 DNA Artificial primer 16gagttattct cgagtttgtc caaatttggc ttagttttac 40 17 30 DNA Artificialprimer 17 tcagtgagat cttgaatacg acacaccatc 30 18 33 DNA Artificialprimer 18 gatttgagtt gtcgacttat ttgtccaaat ttg 33 19 36 DNA Artificialprimer 19 cggagtgcca tatgagctta attaataaat taaatg 36 20 31 DNAArtificial primer 20 tataactcga ggttttgtgc aacaggtgtt g 31 21 34 DNAArtificial primer 21 cgcttgagat cttggaagat gtgtatcagc gtgc 34 22 37 DNAArtificial primer 22 caataacaaa gctttcagtt ttgtgcaaca ggtgttg 37 23 33DNA Artificial primer 23 aaccgcacat atgtatcgct tggtgtcacc acc 33 24 35DNA Artificial primer 24 ggtgactcga ggtactcatc accaactaat cgcac 35 25 27DNA Artificial primer 25 gcaggatcct tatcgcttgg tgtcacc 27 26 31 DNAArtificial primer 26 atcaatcggg tcgacttagt actcatcacc a 31 27 34 DNAArtificial primer 27 aaagcttcat atggcccaaa gtcaagaatc tgcc 34 28 31 DNAArtificial primer 28 cgataactcg agttgaacat caggcacctg c 31 29 39 DNAArtificial primer 29 accattcaaa agagatcttg gcccaaagtc aagaatctg 39 30 33DNA Artificial primer 30 gttagaccga gtcgactcat tgaacatcag gca 33

1-31. (canceled)
 32. An isolated polynucleotide comprising apolynucleotide chosen from: (a) a polynucleotide encoding a polypeptidehaving at least 70% identity to a second polypeptide comprising asequence chosen from: SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments oranalogs thereof; (b) a polynucleotide encoding a polypeptide having atleast 80% identity to a second polypeptide comprising a sequence chosenfrom: SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof;(c) a polynucleotide encoding a polypeptide having at least 95% identityto a second polypeptide comprising a sequence chosen from: SEQ ID NO: 2,4, 6, 8, 10, 12, 14 or fragments or analogs thereof; (d) apolynucleotide encoding a polypeptide comprising a sequence chosen from:SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof; (e) apolynucleotide encoding a polypeptide capable of raising antibodieshaving binding specificity for a polypeptide comprising a sequencechosen from: SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogsthereof; (f) a polynucleotide encoding an epitope bearing portion of apolypeptide comprising a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10,12, 14 or fragments or analogs thereof; (g) a polynucleotide comprisinga sequence chosen from SEQ ID NO: 1, 3, 5, 7, 9, 11, 13 or fragments oranalogs thereof; (h) a polynucleotide that is complementary to apolynucleotide in (a), (b), (c), (d), (e), (f) or (g).
 33. Thepolynucleotide of claim 32, wherein said polynucleotide is DNA.
 34. Thepolynucleotide of claim 32, wherein said polynucleotide is RNA.
 35. Thepolynucleotide of claim 32 that hybridizes under stringent conditions toeither (a) a DNA sequence encoding a polypeptide or (b) the complementof a DNA sequence encoding a polypeptide; wherein said polypeptidecomprises a sequence chosen from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 orfragments or analogs thereof.
 36. The polynucleotide of claim 32 thathybridizes under stringent conditions to either (a) a DNA sequenceencoding a polypeptide or (b) the complement of a DNA sequence encodinga polypeptide; wherein said polypeptide comprises at least 10 contiguousamino acid residues from a polypeptide comprising a sequence chosen fromSEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof.
 37. Avector comprising the polynucleotide of claim 32, wherein said DNA isoperably linked to an expression control region.
 38. A host celltransfected with the vector of claim
 37. 39. A process for producing apolypeptide comprising culturing a host cell according to claim 38 underconditions suitable for expression of said polypeptide.
 40. An isolatedpolypeptide comprising a polypeptide chosen from: (a) a polypeptidehaving at least 70% identity to a second polypeptide having an aminoacid sequence comprising a sequence chosen from: SEQ ID NO: 2, 4, 6, 8,10, 12, 14 or fragments or analogs thereof; (b) a polypeptide having atleast 80% identity to a second polypeptide having an amino acid sequencecomprising a sequence chosen from: SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 orfragments or analogs thereof; (c) a polypeptide having at least 95%identity to a second polypeptide having an amino acid sequencecomprising a sequence chosen from: SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 orfragments or analogs thereof; (d) a polypeptide comprising a sequencechosen from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogsthereof; (e) a polypeptide capable of raising antibodies having bindingspecificity for a polypeptide comprising a sequence chosen from SEQ IDNO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof; (f) anepitope bearing portion of a polypeptide comprising a sequence chosenfrom SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments or analogs thereof;(g) the polypeptide of (a), (b), (c), (d), (e) or (f) wherein theN-terminal Met residue is deleted; (h) the polypeptide of (a), (b), (c),(d), (e), or (f) wherein the secretory amino acid sequence is deleted.41. A chimeric polypeptide comprising two or more polypeptides having asequence chosen from SEQ ID NO: 2, 4, 6, 8, 10, 12, 14 or fragments oranalogs thereof; provided that the polypeptides are linked as to form achimeric polypeptide.
 42. A pharmaceutical composition comprising apolypeptide according to claim 40 and a pharmaceutically acceptablecarrier, diluent or adjuvant.
 43. A method for prophylactic ortherapeutic treatment of Moraxella infection in a host susceptible toMoraxella infection comprising administering to said host a prophylacticor therapeutic amount of a composition according to claim
 42. 44. Amethod according to claim 43 wherein the host is a neonate, an infant ora child.
 45. A method according to claim 43 wherein the host is animmunocompromised host.
 46. A method according to claim 43 wherein thehost is an adult.
 47. A method for therapeutic or prophylactic treatmentof otitis media, sinusitis, persistent cough, acute laryngitis,suppurative keratitis, conjunctivitis neonatorum, and invasive diseasecomprising administering to said host a therapeutic or prophylacticamount of a composition according to claim
 42. 48. A method fordiagnosis of Moraxella infection in a host susceptible to Moraxellainfection comprising (a) obtaining a biological sample from a host; (b)incubating an antibody or fragment thereof reactive with a polypeptideaccording to claim 40 with the biological sample to form a mixture; and(c) detecting specifically bound antibody or bound fragment in themixture which indicates the presence of Moraxella.
 49. A method for thedetection of antibody specific to a Moraxella antigen in a biologicalsample containing or suspected of containing said antibody comprising(a) obtaining a biological sample from a host; (b) incubating one ormore polypeptides according to claim 40 or fragments thereof with thebiological sample to form a mixture; and (c) detecting specificallybound antigen or bound fragment in the mixture which indicates thepresence of antibody specific to Moraxella.
 50. Use of thepharmaceutical composition according to claim 42 in the manufacture of amedicament for the prophylactic or therapeutic treatment of Moraxellainfection.
 51. Kit comprising a polypeptide according to claim 40 fordetection or diagnosis of Moraxella infection.